title: Astrocyte LFS-1 H3K27ac CHIP

data_processing:
QC: FastQC v0.11.5. Sequenced reads were then trimmed for adaptor sequence, and masked for low-complexity or low-quality sequence
RNAeq Alignment: RNAStar for RNASeq, Bowtie2 for ChIPseq
RNAseq normalization: Featurecounts
ChIPseq sorting, normalization, and visualization: MACS2
Genome_build: hg19
Supplementary_files_format_and_content: Supplementary_files_format_and_content: RNASeq: tab-delimited text files include FPKM values for each Sample .csv; MeRIP-seq: .bigwig; ChIPSeq: genome browser tracks in .bigwig

source_name: Astrocytes

growth_protocol: iSPC cell–derived astrocytes cells were routinely generated by passaging 1:3 ratio to adherent astrocytes cultures on matrigel coated plastic in AM medium with AM supplemental, 2% FBS and 10% P/S.

extract_protocol:
For Me-RIPseq, cells samples were lysed in Trizol reagent (Invitrogen, 10296010). mRNA were isolated from total RNA by using mRNA isolation kit (11741985001 Roche). mRNA were fragmented by 10 mM Zncl2 solution and incubated with protein A/G dynabeas and equal amount of m6A antibody (SYSY) to mRNA. Input mRNA were not with antibody incubation. After wash, RNA were extracted by using TrizOL
For RNAseq, cells samples were lysed in Trizol reagent (Invitrogen, 10296010).ChIP-seq, cells were fixed, quenched, sonicated to ~250 bp in 1% SDS lysis buffer, and incubated with Protein A/G Dynabeads and 5 µg of the indicated antibody. 2% pre-cleared chromatin prior to primary antibody addition was kept as input DNA. After wash, treatment with RNAse A and Proteinase K, DNA was eluted and purified with Qiagen's PCR purification kit (28106).
For RNASeq, library construction was performed using NEBNext Ultra II RNA Library Prep Kit (E7770) following manufacturer’s instructions. For ChIPSeq, library construction was performed using KAPA Hyper Prep Kit (KK8500) according to manufacturer's conditions. the poly(A) selection step and starting with the RNA fragmentation step.Libraries were submitted to PE150 high-throughput sequencing.

treatment_protocol: Treated with OICR-9429 8uM for 2 days

tag: chip antibody

value: H3K27ac (Abcam, ab4729)

value: LFS Patient


title: Old adult CBA/CaJ mouse LW-3

status: Public on Feb 18 2023

type: SRA

city: Charleston

first: Jeremy

last: Barth

state: SC

address: 173 Ashley Ave, BSB 627

country: USA

institute: MUSC

library_source: transcriptomic

data_processing:
Illumina Casava1.8 software used for basecalling.
Sequenced reads (fastq files) were trimmed for adaptor sequence, and masked for low-complexity or low-quality sequence
Secondary analysis was carried out using Partek Flow. Alignment was conducted with TopHat2 using mouse genome (mm10) and the default option set
Aligned reads were quantified to annotation model (Partek E/M) using mm10 and RefSeq transcripts 83 (2017-11-01) with the settings: a) strict paired-end compatibility, b) require junction reads to match introns, c) min read overlap with feature 100% of read length, d) min reads 10
Gene counts were analyzed using DESeq2 to determine normalized expression measures and adjusted p-value (FDR step up) for old versus young samples
Genome_build: mm10
Supplementary_files_format_and_content: tab-delimited text file of normalized counts and comparison measures

molecule: total RNA

organism: Mus musculus

source_name: Auditory nerve cells

extract_protocol:
Microdissections were performed to isolate the cochle lateral wall from mouse cochlear structures. Two cochleas from each mouse were pooled for individual samples. Total RNA was purified from each sample using the miRNeasy Mini Kit per the manufacturer’s instructions. Quality of each total RNA isolation was assessed using the Agilent 2100 Bioanalyzer and low-quality samples showing degradation or contamination were excluded.
RNA libraries were prepared for sequencing using standard Illumina protocols using Illumina's TruSeq RNA Library Prep Kit

tag: tissue

value: Isolated lateral wall

tag: age

tag: strain

value: CBA/CaJ


title: Astrocyte WT-1 H3K27ac CHIP

city: Houston

first: An

last: Xu

state: TX

institute: UThealth Houston

library_source: genomic

molecule: genomic DNA

organism: Homo sapiens

tag: cell type

value: iPSC-derived astrocytes

tag: treatment

value: No

tag: genotype

value: Healthy Family Control

tag: disease state

value: Normal


title: GEM23

city: Pullman

first: Michael

middle: K

last: Skinner

state: WA

address: Abelson 507

department: SBS

institute: WSU

description:
female.all.results.csv.gz
female.homo.results.csv.gz

data_processing:
Basic read quality was verified using summaries produced by FastQC. Trimmomatic was used to remove low quality reads and bases
The reads for each sample were mapped to the human (GRCh38) genome using Bowtie2
The mapped read files were then converted to sorted BAM files using SAMtools
The MEDIPS R package was used to calculate differential coverage between sample groups. To identify DMR, the reference genome was broken into 1000 bp windows. The edgeR p-value was used to determine the relative difference between the two sample groups for each genomic window. Windows with an edgeR p-value less than an arbitrarily chosen threshold were considered DMR. The DMR edges were extended until no genomic window with an edgeR p-value less than 0.1 remained within 1000 bp of the DMR.
The selected DMR were then summarized and annotated.
Assembly: GRCh38
Supplementary files format and content: The results of the edgeR analysis in CSV format. This includes raw read counts for each genomic window and other summary values.

source_name: buccal

growth_protocol: Participants for this study were MZ twins recruited from the community-based Washington State Twin Registry (WSTR). Participants in the current study previously participated in a study of objective measures of physical activity (PA) and locations among individuals residing in the Puget Sound area around Seattle between 2012 and 2015. Follow-up data collection was conducted in 2018 and 2019, 72 pairs completed follow-up collection out of the 144 pairs who completed the baseline study.

extract_protocol:
Frozen human buccal samples were thawed for analysis. Genomic DNA from buccal samples was prepared as follows: The buccal brush was suspended in 750 μl of cell lysis solution and 3.5 µl of Proteinase K (20 mg/ml). This suspension was incubated at 55ºC for 3 hours, then vortexed and centrifuged briefly. The lysis solution was then transferred to a new 1.5 µl microcentrifuge tube. The microcentrifuge tube with the buccal brush was centrifuged again to retain any remaining solution which was combined with the transferred lysis solution. The buccal brush was discarded and 300 µl of protein precipitation solution (Promega, A795A, Madison, WI) was added to the lysis solution. The sample was incubated on ice for 15 minutes, then centrifuged at 4C for 30 minutes. The supernatant was transferred to a fresh 2 mL microcentrifuge tube and 1,000 µl ice cold isopropanol was added along with 2 µL glycoblue. This suspension was mixed thoroughly and incubated at -20 ºC overnight. The suspension was then centrifuged at 4ºC for 20 minutes, the supernatant was discarded, and the pellet was washed with 75% ethanol, then air-dried and resuspended in 100 μl H2O. DNA concentration was measured using the Nanodrop (Thermo Fisher, Waltham, MA). Methylated DNA Immunoprecipitation (MeDIP) with genomic DNA was performed as follows: individual DNA samples (2-4 ug of total DNA) were diluted to 130 μl with 1x Tris-EDTA (TE, 10 mM Tris, 1 mM EDTA) and sonicated with the Covaris M220 using the 300 bp setting. Fragment size was verified on a 2% E-gel agarose gel. The sonicated DNA was transferred from the Covaris tube to a 1.7 ml microfuge tube, and the volume was measured. The sonicated DNA was then diluted with TE buffer (10mM Tris HCl, pH7.5; 1mM EDTA) to 400 μl, heat-denatured for 10 min at 95 C, then immediately cooled on ice for 10 min. Then 100 μl of 5X IP buffer and 5 μg of antibody (monoclonal mouse anti 5-methyl cytidine; Diagenode #C15200006) were added to the denatured sonicated DNA. The DNA-antibody mixture was incubated overnight on a rotator at 4 C. The following day magnetic beads (Dynabeads M-280 Sheep anti-Mouse IgG; 11201D) were pre-washed as follows: The beads were resuspended in the vial, then the appropriate volume (50 μl per sample) was transferred to a microfuge tube. The same volume of Washing Buffer (at least 1 mL 1XPBS with 0.1% BSA and 2mM EDTA) was added and the bead sample was resuspended. The tube was then placed into a magnetic rack for 1-2 min and the supernatant was discarded. The tube was removed from the magnetic rack and the beads were washed once. The washed beads were resuspended in the same volume of 1xIP buffer (50 mM sodium phosphate ph7.0, 700 mM NaCl, 0.25% TritonX-100) as the initial volume of beads. 50μl of beads were added to the 500μl of DNA-antibody mixture from the overnight incubation, then incubated for 2 hours on a rotator at 4 C. After the incubation, the bead-antibody-DNA complex was washed three times with 1X IP buffer as follows: The tube was placed into a magnetic rack for 1-2 min and the supernatant was discarded, then the magnetic bead antibody pellet was washed with 1xIP buffer 3 times. The washed bead antibody DNA pellet was then resuspended in 250 μl digestion buffer with 3.5 μl Proteinase K (20mg/ml). The sample was incubated for 2-3 hours on a rotator at 55 C, then 250 μl of buffered Phenol-Chloroform- Isoamylalcohol solution was added to the sample, and the tube was vortexed for 30 sec and then centrifuged at 14,000rpm for 5 min at room temperature. The aqueous supernatant was carefully removed and transferred to a fresh microfuge tube. Then 250 μl chloroform were added to the supernatant from the previous step, vortexed for 30 sec and centrifuged at 14,000rpm for 5 min at room temperature. The aqueous supernatant was removed and transferred to a fresh microfuge tube. To the supernatant 2μl of glycoblue (20mg/ml), 20μl of 5M NaCl and 500μl ethanol were added and mixed well, then precipitated in -20 C freezer for 1 hour to overnight. The precipitate was centrifuged at 14,000rpm for 20 min at 4 C and the supernatant was removed, while not disturbing the pellet. The pellet was washed with 500μl cold 70% ethanol in -20 C freezer for 15 min then centrifuged again at 14,000rpm for 5 min at 4 C and the supernatant was discarded. The tube was spun again briefly to collect residual ethanol to the bottom of the tube and as much liquid as possible was removed with gel loading tip. The pellet was air-dried at RT until it looked dry (about 5 min) then resuspended in 20μl TE. DNA concentration was measured in Qubit (Life Technologies) with ssDNA kit (Molecular Probes Q10212).
The MeDIP DNA samples (50 ng of each) were used to create libraries for next generation sequencing (NGS) using the NEBNext Ultra RNA Library Prep Kit for Illumina (San Diego, CA) starting at step 1.4 of the manufacturer’s protocol to generate double stranded DNA. After this step the manufacturer’s protocol was followed. Each sample received a separate index primer.

treatment_protocol: Once participants were enrolled in the study, the study coordinator sent all study materials to the participant for remote data collection. Participants wore a Qstarz BT-Q1000XT (Qstarz International Co. Ltd, Taipei, Taiwan) GPS data logger and Actigraph GT3X+ accelerometer (Actigraph Inc. Pensacola, FL) attached to a belt worn around the waist for one week. They also completed questionnaires and provided a buccal sample, with 70 complete pairs provided buccal samples. The buccal cell collection procedure involved 30 second brushing of each cheek before putting in enclosed vial for shipment by mail and then stored at -80°C. Materials were sent to the Skinner lab at WSU Pullman after sample and data collection was completed. Buccal swab brushes were stored at -80°C until use. The study protocol was approved by the Washington State University Institutional Review Board (#16419), and informed written consent was obtained from all participants prior to receiving the study materials. All methods were carried out in accordance with relevant guidelines and regulations.

tag: twin pair_number

tag: srzyg

value: homozygote

tag: Sex

value: female

value: buccal

tag: average waist

tag: bmi

tag: obese call

value: non-obese


title: Astrocyte LFS-1 p53 CHIP

value: p53 (Santa Cruz  Technology,sc-126)

value: LFS


title: Astrocyte WT-1 p53 CHIP


title: GEM28

value: obese (I)


title: GEM21

description:
male.all.results.csv.gz
male.homo.results.csv.gz

value: male


title: GEM27


title: MFB cells after a 24h of non-contact co-culture with umbilical cord blood-derived mesenchymal stem cells (uMSC), #1

city: Guangzhou

first: Ruohao

state: Guangdong

address: Zhongshan Second Road, Guangzhou, China

department: Hematology

country: China

institute: Guangdong Provincial People's Hospital

data_processing:
CLC Genomics Workbench v 11.0.1
Sequence reads were trimmed for adaptor sequence/low-quality sequence using CLC genomic benchwork (parameter- Quality limit: 0.05)
Trimmed sequence reads were mapped to GrCM38/mm10 using CLC genomic benchwork (parameters- mismath cost: 2 insertion cost: 3 deletion cost: 3 length fraction: 0.8 similarity fraction: 0.8)
Read count extraction and normalization were performed using CLC genomic benchwork
Assembly: GRCh38
Supplementary files format and content: Raw readcounts for each sample
Supplementary files format and content: Normalized FPKM value for each sample

source_name: lung

growth_protocol: FB cells were cultured using DMEM supplemented with 10% fetal bovine serum (FBS) containing 100 units/mL penicillin and 100 ug/mL streptomycin (both from Gibco)

extract_protocol:
Briefly, mRNA was purified from total RNA using poly-T oligo-attached magnetic   beads. Fragmentation was carried out using divalent cations under elevated temperature   in NEBNext First Strand Synthesis Reaction Buffer(5X). First strand cDNA was   synthesized using random hexamer primer and M-MuLV Reverse
Sequencing   libraries were generated using NEBNext® UltraTM RNA Library Prep Kit for   Illumina® (NEB, USA) following manufacturer’s recommendations and index codes   were added to attribute sequences to each sample.

treatment_protocol: For MFB differentiation, FB was serum-starved in FBS-free DMEM overnight and stimulated with 10ng/mL TGF-β1 (PeroTech, USA) for 48h to induce MFB differentiation

value: lung

tag: cell line

value: FB-like cell

value: WT


title: MFB cells (FB cells after a 48h treatment of single-dose TGF-β1 at 10ng/mL), sample #2

value: myofibroblast

value: 48h treatment of single-dose TGF-Beta1 at 10ng/mL


title: MFB cells (FB cells after a 48h treatment of single-dose TGF-β1 at 10ng/mL), sample #1


title: Untreated MRC-5 human lung fibroblast cells, sample #2

value: fibroblast

value: DMSO


title: GEM20

description:
female.all.results.csv.gz
female.hetero.results.csv.gz

value: heterozygote


title: Untreated MRC-5 human lung fibroblast cells, sample #1


title: GEM22


title: Astrocytes LFS with OICR-9429

value: iPSC-derived astrocytes

value: Yes (OICR-9429 8uM for 2 days)

value: H3K4me3 (Abcam, ab8580)

value: LFS Patient with treatment


title: Astrocytes LFS DMSO

value: No

value: LFS Patient with DMSO


title: H446_shNFIB_Input_rep2

city: Charlestown

first: Rui

last: Dong

state: MA

address: 149 13th Street

institute: Massachusetts General Hospital

data_processing:
Illumina Casava v.2.19 software used for basecalling.
Sequenced reads were aligned to hg19 genome using bwa with default settings.
Duplicate reads were removed using picard-tools v.1.95, then gap regions within paired reads were filled.
Assembly: hg19
Supplementary files format and content: bigwig files were generated using bedtools; scores show fragments normalized to 10M mapped fragments.
Library strategy: DisP-seq

source_name: small cell lung cancer cells

growth_protocol: NCI-H446 was grown in specific RPMI 1640 Medium (Gibco, A1049101) and SKNMC was grown in RPMI-1640 (Gibco, 11875). MRC5 was grown in EMEM (ATCC, 30-2003). All media were supplemented with 10% FBS and cells were cultured at 37°C with 5% CO2.

extract_protocol:
Lysates were clarified from Mnase digested nuclei and protein-DNA complexes were isolated with biotinylated isoxazole.
Precipitated DNA samples were used to prepare sequencing libraries with Ultralow V2 DNA-Seq Library Preparation Kit (NuGEN, 0344NB-A01), and DNA samples were sequenced with the Nextseq 500 Illumina genome analyzer.

treatment_protocol: untreated

value: small cell lung cancer cells

value: untreated


title: GEM18

value: female

value: buccal

value: obese (III)


title: SKNMC, shGFP - Input, rep2

data_processing:
Library strategy: DisP-seq
Illumina Casava v.2.19 software used for basecalling.
Sequenced reads were aligned to hg19 genome using bwa with default settings.
Duplicate reads were removed using picard-tools v.1.95, then gap regions within paired reads were filled.
Genome_build: hg19
Supplementary_files_format_and_content: bigwig files were generated using bedtools; scores show fragments normalized to 10M mapped fragments.

source_name: Ewing sarcoma cells

value: Ewing sarcoma cells

value: SKNMC

value: shGFP control

tag: precipitation reagent


title: MFB cells (FB cells after a 48h treatment of single-dose TGF-β1 at 10ng/mL), sample #3

value: lung

value: myofibroblast

value: WT

value: 48h treatment of single-dose TGF-Beta1 at 10ng/mL


title: Astrocytes WT with OICR-9429

value: iPSC-derived astrocytes

value: Yes (OICR-9429 8uM for 2 days)

value: Healthy Family Control with treatment

value: Normal


title: SKNMC, shGFP, DMSO, rep2

value: DMSO


title: SKNMC, shGFP - Input, rep1


title: Astrocytes WT DMSO

value: No

value: H3K4me3 (Abcam, ab8580)

value: Healthy Family Control with DMSO


title: Astrocyte LFS-MLL1 KD

value: LFS Patient with knockdown

value: LFS


title: MFB cells after a 24h of non-contact co-culture with umbilical cord blood-derived mesenchymal stem cells (uMSC), #3

value: FB-like cell


title: SKNMC, shGFP, DMSO, rep1

value: Ewing sarcoma cells

value: SKNMC

value: untreated


title: GEM16

value: homozygote

value: female

value: buccal

value: obese (I)


title: Untreated MRC-5 human lung fibroblast cells, sample #3

value: lung

value: fibroblast

value: WT


title: GEM19

value: heterozygote

value: non-obese


title: SKNMC, shGFP, bisox, rep1

value: shGFP control

value: b-isox


title: GEM15


title: SKNMC, shFLI1 - Input, rep2

value: EWS-FLI1 knockdown


title: Astrocyte LFS m6A

description: mRNA IP with m6A

value: iPSC-derived astrocytes

value: Yes (OICR-9429 8uM for 2 days)

tag: rip antibody

value: m6A (Synaptic System, 202-003)

value: LFS Patient with m6A pull down

value: LFS


title: SKNMC, shFLI1 - Input, rep1

value: Ewing sarcoma cells

value: SKNMC

value: untreated


title: Astrocyte LFS-YTHDF2 KD2

description: RNAseq FPKM3.csv

value: LFS Patient with knockdown


title: Astrocyte LFS m6A input

description: mRNA

value: No

value: LFS Patient


title: GEM14

value: female

value: buccal

value: obese (I)


title: GEM17

value: heterozygote

value: non-obese


title: SKNMC, shFLI1, DMSO, rep2

value: EWS-FLI1 knockdown

value: DMSO


title: SKNMC, shFLI1, DMSO, rep1


title: MFB cells after a 24h of non-contact co-culture with umbilical cord blood-derived mesenchymal stem cells (uMSC), #2

value: lung

value: FB-like cell

value: WT


title: SKNMC, shFLI1, bisox, rep2

value: b-isox


title: Astrocyte LFS-YTHDF2 KD1

description: RNAseq FPKM3.csv

value: iPSC-derived astrocytes

value: Yes (OICR-9429 8uM for 2 days)

value: LFS


title: SKNMC, shFLI1, bisox, rep1

value: Ewing sarcoma cells

value: SKNMC

value: untreated


title: GEM13

value: female


title: SKNMC - Input, rep1

value: wildtype


title: SKNMC, DMSO, rep2


title: SKNMC - Input, rep2


title: Astrocyte LFS-ctrl1

description: RNAseq FPKM3.csv

value: No

value: LFS Patient Scramble


title: Astrocyte LFS-OICR-9429-2

description: RNAseq FPKM2.csv

value: LFS Patient with treatment


title: SKNMC, bisox, rep2


title: SKNMC, shGFP, bisox, rep2

value: shGFP control


title: GEM12

value: homozygote

value: buccal

value: obese (I)


title: Astrocyte  LFS-OICR-9429-1

description: RNAseq FPKM2.csv


title: GEM10


title: SKNMC, bisox, rep1


title: SKNMC, DMSO, rep1

value: DMSO


title: GEM9

value: non-obese


title: SKNMC, 6U, bisox

value: Nucleus digestion with 12 U Mnase (MNase titration of DisP-seq to test the efficiency of DNA digestion)

value: b-isox


title: Astrocyte LFS-ctrl2 DMSO

description: RNAseq FPKM2.csv

value: iPSC-derived astrocytes

value: LFS Patient with DMSO

value: LFS


title: GEM8

value: obese (II)


title: SKNMC, 12U, bisox

value: Ewing sarcoma cells

value: SKNMC


title: GEM6

value: heterozygote

value: female


title: Astrocyte LFS-CDKN2B OV2

description: RNAseq FPKM2.csv

value: Yes (OICR-9429 8uM for 2 days)

value: LFS Patient restored CDKN2B


title: SKNMC, 3U, bisox

value: wildtype


title: GEM11

value: homozygote

value: buccal


title: MRC5, DMSO, rep2

source_name: fibroblast cells

value: fibroblast cells

value: wild type

value: untreated

value: DMSO


title: Astrocyte LFS-CDKN2B OV1

description: RNAseq FPKM2.csv

value: iPSC-derived astrocytes


title: Astrocyte LFS-ctrl1 DMSO

description: RNAseq FPKM2.csv

value: No

value: LFS Patient with DMSO


title: MRC5, DMSO, rep1


title: GEM5

value: non-obese


title: Astrocyte LFS-ctrl2 ov

description: RNAseq FPKM2.csv

value: LFS Patient

value: LFS


title: MRC5, bisox, rep1

value: b-isox


title: Astrocyte LFS-ctrl1 ov

description: RNAseq FPKM2.csv


title: H446, shNFIB - Input

value: small cell lung cancer cells

value: NFIB knockdown


title: GEM4

value: female

value: obese (I)


title: H446, shNFIB, DMSO, rep2


title: H446, shNFIB, DMSO, rep1


title: GEM3

value: homozygote


title: H446, shNFIB, bisox, rep2


title: Astrocyte H1-245-2

description: RNAseq FPKM1.csv

value: hES-derived astrocytes

value: Yes (OICR-9429 8uM for 2 days)

value: hES with p53 mutation


title: Astrocyte H1-245-1

description: RNAseq FPKM1.csv


title: MRC5 - Input, rep1

value: fibroblast cells

value: wild type

value: untreated


title: MRC5 - Input, rep2


title: GEM7

value: buccal


title: SKNMC, 24U, bisox

value: Ewing sarcoma cells

value: SKNMC

value: wildtype

value: Nucleus digestion with 12 U Mnase (MNase titration of DisP-seq to test the efficiency of DNA digestion)


title: GEM2

value: heterozygote

value: obese (III)


title: GEM1

value: non-obese


title: H446, shNFIB, bisox, rep1

value: small cell lung cancer cells

value: NFIB knockdown

value: b-isox


title: MRC5, bisox, rep2


title: H446, shGFP - Input, rep2

value: shGFP control


title: Astrocyte H1-3

description: RNAseq FPKM1.csv

value: hES-derived astrocytes

value: No

value: Normal hES Control

value: Normal


title: H446, shGFP - Input, rep1

value: untreated


title: HSC_midage_LEPRneg_rep7

city: Indianapolis

first: James

last: Ropa

state: IN

address: 950 W Walnut St

institute: Indiana University

data_processing:
Quality of reads were analyzed with FastQC.
Reads were adapter trimmed and trimmed for quality with Cutadapt with the options -a AGATCGGAAGAGCACACGTC -A AGATCGGAAGAGCGTCGTGT --nextseq-trim=20 --minimum-length 50
Genome indices were generated using the GRCm39 mouse genome downloaded from GENCODE (Release M27) using STAR sequence aligner with the options --sjdbOverhang 99.
Reads were aligned using the user-built genome indices with STAR sequence aligner with the options --readFilesCommand zcat --outSAMtype BAM SortedByCoordinate --outSAMunmapped Within
Reads were counted using HTSeq assigning reads to genes using the options -s no -r pos -f bam
Reads were normalized based on library size and differential expression was assessed using DESeq2
Assembly: mm39
Supplementary files format and content: counts files: space delimited files containing unnormalized read counts assigned to genes generated by HTSeq.

molecule: polyA RNA

source_name: bone marrow

extract_protocol:
Bone marrow from young (2-3 months) or middle-aged (12-15months) was harvested by flushing femurs and tibiae and performing a affinity based magnetic lineage depletion. HSCs (Lin-Sca1+Kit+CD150+CD48-) were sorted using flow cytometry by LEPR status (LEPR+ HSCS and LEPR- HSCs). RNA was harvested using Qiagen Rneasy Micro Plus Kit.
NEBNext Single Cell/ Ultra Low Input RNA-seq library prep kit was used to prepare libraries per manufacturer recommendations.

value: bone marrow

value: Hematopoietic stem cells

value: female

tag: phenotype

value: LEPR negative

tag: mouse number

tag: developmental stage

value: midage


title: HSC_midage_LEPRneg_rep6

tag: tissue


title: Astrocyte H1-1

description: RNAseq FPKM1.csv

tag: cell type


title: Astrocyte LFS-2

description: RNAseq FPKM1.csv

value: iPSC-derived astrocytes

tag: treatment

value: Yes (OICR-9429 8uM for 2 days)

value: LFS Patient

value: LFS


title: Astrocyte H1-2

description: RNAseq FPKM1.csv

tag: genotype


title: H446, shGFP, DMSO, rep1

value: small cell lung cancer cells

value: DMSO


title: Flt_CD8_2

city: Wuhan

first: Hong-Peng

state: Hubei

address: Donghu Road, No. 115, Wuchang District

institute: Wuhan University

data_processing:
Sequence reads were trimmed for adaptor sequence/low-quality sequence using Trimmomatic (version 0.36) (parameter- ILLUMINACLIP:{adaptor.fa}:2:30:10  -phred33 LEADING:3 TRAILING:3 SLIDINGWINDOW:4:15 HEADCROP:0 MINLEN:36 PE)
Clean Reads were further treated with UMI_soft (in-house) to eliminate duplication bias introduced in library preparation and sequencing on using UMI. （Default Parameter developed by Wuhan Seqhealth Co., Ltd.）
The de-duplicated consensus sequences were mapped to GRCh38 using STAR software (version 2.5.3a) (parameters- --outSAMtype BAM SortedByCoordinate)
Read count extraction and normalization were performed using featureCounts（Version 1.5.1）（parameter -T 10 -d 30 -D 1000 -C -s 1 -t {exon} -g {geneid} --primary -O -a {gff_annotation_file}）
Assembly: Mus_musculus.GRCm38
Supplementary files format and content: tab-delimited text files include RPKM values for each Sample

growth_protocol: CD8+ T cells sorted from lung tumors of KP mice that were intranasally injected with Ad-Cre (1.5x106 pfu per mouse)

extract_protocol:
RNA was harvested using TRIzol Reagent (Invitrogen, cat. NO 15596026）. 2 ug of total RNA was used for the construction of sequencing libraries.
RNA libraries for RNA-seq were prepared using KC-DigitalTM Stranded mRNA Library Prep Kit for Illumina® (Catalog NO. DR08502, Wuhan Seqhealth Co., Ltd. China) following manufacturer's protocols.

treatment_protocol: KP mice intraperitoneal injection with PBS, PD-1Ab,PD-1Ab7 , or PD-1Ab7 plus Flt3L

value: lung

value: CD8+ T cell

value: WT

value: Flt3L plus PD-1Ab7


title: PBS_CD8_2

value: PBS


title: PD1_CD8_2


title: P7_CD8_2


title: HSC_midage_LEPRneg_rep5

value: male


title: HSC_midage_LEPRneg_rep3

value: bone marrow


title: HSC_midage_LEPRneg_rep4

value: Hematopoietic stem cells


title: HSC_midage_LEPRneg_rep2

value: female


title: Astrocyte LFS-1

description: RNAseq FPKM1.csv

tag: disease state


title: H446, shGFP, bisox, rep2

value: shGFP control

value: untreated

value: b-isox


title: PBS_CD8_1


title: Astrocyte WT-2

description: RNAseq FPKM1.csv

value: iPSC-derived astrocytes

value: No

value: Healthy Family Control

value: Normal


title: PD1_CD8_1


title: H446, shGFP, bisox, rep1

value: small cell lung cancer cells


title: HSC_midage_LEPRpos_rep7

value: LEPR positive

value: midage


title: Astrocyte WT-1

description: RNAseq FPKM1.csv


title: HSC_midage_LEPRneg_rep1

value: LEPR negative


title: H446 - Input, rep2

value: wildtype


title: H446, shGFP, DMSO, rep2

value: DMSO


title: H446 - Input, rep1


title: HSC_midage_LEPRpos_rep6


title: brain tumor cells, 5783-TVA-B

city: New York

first: Dolores

last: Hambardzumyan

state: NY

address: 1425 Madison Ave.

department: Oncological Sciences

institute: Icahn School of Medicine

description: 10x Genomics

library_source: transcriptomic single cell

data_processing:
Raw fastq files were aligned to mouse genome reference mm10 customized to include the Rfp sequence, using CellRanger v5.0.0 (10X Genomics). Count matrices filtered by CellRanges algorithm were further filtered by discarding cells with either < 400 genes, < 1000 UMI (unique molecular identifier), or > 25% mitochondrial genes expressed. Data was processed and analyzed using R package Seurat v4.0.5. Normalization was performed using NormalizeData function with normalization.method = ‘LogNormalize’. Dimensionality reduction was computed on the top 2,000 variable features using FindVariableFeatures, ScaleData and RunPCA functions. UMAPs were generated using the top 15 PCs.
For subclustering the immune compartment, we used R package Harmony to mitigate for batch effects driven by technical variation between replicates. De novo clustering using the Louvain algorithm was applied at different resolutions (0.2; 0.8; 2; 5) on the KNN graph space. For high-level annotation, cell types were identified in an iterative and semi-supervised fashion by assigning de novo discovered clusters to cell types based on expression of known marker genes that define each cluster. Annotation of cell subtypes at a lower-level was performed in a similar manner as for the high-level and further aided by de novo marker discovery using the Seurat FindMarkers function and Wilcoxon Rank Sum test.
To identify doublet-enriched clusters we looked for clusters of cells displaying expression of canonical markers for two or more different cell types and higher number of genes/UMI; such clusters were removed from further analysis.
Cell-level proliferation analysis was carried out with Seurat function CellCycleScoring using a list of murine cell cycle genes. Cells were then assigned as ‘cycling’ or ‘non-cycling’ if either their S-phase score or G2M-score was above a threshold of 0.05 for the tumor compartment and 0.1 for the immune compartment.
Identification of modules of co-expressed genes was carried out using the R package scWGCNA (https://github.com/smorabit/scWGCNA)  by first computing metacells of 100 cells (k=100) using function construct_metacells. To identify modules, function blockwiseConsensusModules was called with following parameters: softPower=12, deepSplit=3, mergeCutHeight = 0.25. Only the top 2,000 variable genes were used.
Assembly: mm10
Supplementary files format and content: Cell Ranger output files (barcodes.tsv, features.tsv, matrix.mtx.gz)

source_name: brain tumor

extract_protocol:
Tumors were dissected from the brain, minced into pieces < 1 mm3, and digested with an enzymatic mixture that includes papain (0.94 mg/ml, Worthington, LS003120), EDTA (0.18 mg/ml, Sigma, E6758), cysteine (0.18 mg/ml, Sigma, A8199), and DNase I (60 μg/ml, Roche, 11284932001) in 2 ml HBSS (Gibco, 14175-095). Tumor tissues were kept at 37°C for 15 minutes with occasional agitation. The digestion was terminated with the addition of 2 ml Ovomucoid (0.7 mg/ml, Worthington, LS003086). Following digestion, single cells were passed through a 70 μm cell strainer, centrifuged, and resuspended in 30% Percoll (GE Healthcare, 17-0891-01) solution containing 10% FBS (Hyclone SH30396.03). Cells were separated by centrifugation at 800g for 15 minutes at 4°C. The pellet was resuspended in RBC lysis buffer (BioLegend, 420301) for 1 min at 37°C before they were centrifuged at 450g for 5 min. The pellet was resuspend in PBS and immediately dilvered for GEM preparation.
Gel-Bead in Emulsions (GEMs) were generated on the sample chip in the Chromium controller. Barcoded cDNA was extracted from the GEMs by Post-GEM RT-cleanup and amplified for 12 cycles. Amplified cDNA was fragmented and subjected to end-repair, poly A-tailing, adapter ligation, and 10X-specific sample indexing following the manufacturer’s protocol. Libraries were quantified using Bioanalyzer (Agilent) and QuBit (ThermoFisher) analyses

value: brain tumor cells

value: brain tumor

value: WT;Ntv-a

tag: age

value: 8-16 weeks old


title: HSC_midage_LEPRpos_rep5

value: bone marrow

value: Hematopoietic stem cells

value: male


title: Flt_CD8_1

value: lung

value: CD8+ T cell

value: WT

value: Flt3L plus PD-1Ab7


title: HSC_midage_LEPRpos_rep4


title: H446, DMSO, rep2

value: small cell lung cancer cells

value: untreated


title: H446, DMSO, rep1


title: H446, bisox, rep2

value: b-isox


title: HSC_midage_LEPRpos_rep2

value: female

value: LEPR positive

value: midage


title: HSC_midage_LEPRpos_rep3


title: brain tumor cells, 5726-TVA-B

description: 10x Genomics

value: brain tumor cells


title: Flt_DC

growth_protocol: cDC1s sorted from lung tumors of KP mice that were intranasally injected with Ad-Cre (1.5x106 pfu per mouse)

value: Dendritic Cell


title: brain tumor cells, 5310-WT-B-p53

description: 10x Genomics

value: brain tumor

value: WT;Ntv-a


title: PBS_DC

value: PBS


title: PD1_DC


title: brain tumor cells, 5326-MCP-B-p53-mm10

description: 10x Genomics

value: qMCP;Ntv-a

value: 8-16 weeks old


title: P7_DC

value: lung


title: brain tumor cells, 5311-MCP-B-p53-mm10

description: 10x Genomics


title: P7_CD8_1

value: CD8+ T cell


title: 10144, hippocampus, 16p11_2_Del5

city: ILLKIRCH

first: Yann

last: HERAULT

address: 1 rue Laurent Fries

country: France

institute: IGBMC/PHENOMIN-ICS

data_processing:
Image analysis and base calling were performed using RTA 2.7.3 and bcl2fastq 2.17.1.14.
The raw sequenced reads were aligned by Hisat2 against the Rno6.v96. 32623 ENSEMBL Gene Ids were quantified aligning to the Rno6.v96 assembly. HTSeq-count was used to generate the raw counts.
The downstream analyses were carried on with in-house bash scripts and R version 3.6 scripts using FCROS and DESeq2 packages to identify the DEGs. Raw reads and normalized counts have been deposited in GEO
Assembly: rn6
Supplementary files format and content: raw counts

organism: Rattus norvegicus

source_name: Hippocampus

extract_protocol: RNA-Seq libraries were generated from 250 ng of total RNA using TruSeq Stranded mRNA Library Prep Kit and TruSeq RNA Single Indexes kits A and B (Illumina, San Diego, CA), according to manufacturer's instructions. Briefly, following purification with poly-T oligo attached magnetic beads, the mRNA was fragmented using divalent cations at 94oC for 2 minutes. The cleaved RNA fragments were copied into first strand cDNA using reverse transcriptase and random primers. Strand specificity was achieved by replacing dTTP with dUTP during second strand cDNA synthesis using DNA Polymerase I and RNase H. Following addition of a single 'A' base and subsequent ligation of the adapter on double stranded cDNA fragments, the products were purified and enriched with PCR (30 sec at 98oC; [10 sec at 98oC, 30 sec at 60oC, 30 sec at 72oC] x 12 cycles; 5 min at 72oC) to create the cDNA library. Surplus PCR primers were further removed by purification using AMPure XP beads (Beckman-Coulter, Villepinte, France) and the final cDNA libraries were checked for quality and quantified using capillary electrophoresis.

tag: genetic background

value: Long Evans

value: male

value: Hippocampus


title: X88_p7

city: Taipei City

first: Chi-Yuan

last: Yao

address: No.7, Chung Shan S. Rd.

country: Taiwan

institute: National Taiwan University Hospital

data_processing:
Sequenced reads were aligned to mouse genome GRCm38 using TopHat v2.1.1.
Gene-level expression was quantified by htseq-count (version 0.11.2) based on GENCODE annotation version 6 (Ensembl 81).
Read counts were then normalized by DESeq.
Genome_build: GRCm38
Supplementary_files_format_and_content: Tab-delimited text file of DESeq normalized read counts

extract_protocol:
Mouse bone marrow cells were harvested from young Hopx-wildtype and Hopx-knockout mice, and sorted into subpopulations using BD FACSAriaIII multicolor cell sorter
RNA libraries were prepared for sequencing using Nextera XT DNA sample preparation kit (Illumina, San Diego, CA)

tag: strain background

value: 8 weeks

tag: genotype/variation

value: Hopx-knockout

value: Bone marrow

value: type 2 multipotent progenitors (MPP2)


title: 10142, hippocampus, 16p11_2_Del4

tag: Sex


title: brain tumor cells, 5303-MCP-B-53-mm10

description: 10x Genomics


title: HSC_midage_LEPRpos_rep1

value: bone marrow

value: Hematopoietic stem cells

value: female

value: LEPR positive

value: midage


title: HSC_young_LEPRneg_rep4

value: LEPR negative

value: young


title: 10125, hippocampus, 16p11_2_Del3


title: HSC_young_LEPRneg_rep2


title: X87_p6

value: type 3 multipotent progenitors (MPP3)


title: HSC_young_LEPRneg_rep3


title: X86_p8

value: Hopx-wildtype

value: long-term HSC (LT-HSCs)


title: HSC_young_LEPRneg_rep1


title: H446, bisox, rep1

value: small cell lung cancer cells

value: wildtype

value: untreated

value: b-isox


title: X84_p6

value: 8 weeks


title: 10109, hippocampus, 16p11_2_Del1

value: Long Evans

value: male

value: Hippocampus


title: HSC_young_LEPRpos_rep4

value: bone marrow

value: Hematopoietic stem cells


title: HSC_young_LEPRpos_rep3

value: female

value: LEPR positive


title: 10107, hippocampus, 16p11_2_wt5

value: wt


title: X81_p6

value: Bone marrow


title: Celf1_KO_P6_A

type: RNA

city: Newark

first: Salil

middle: Anil

last: Lachke

state: Delaware

address: 351 Wolf Hall, University of Delaware

department: Biological Sciences

institute: University of Delaware

hyb_protocol: Standard Illumina hybridization protocol

scan_protocol: Standard Illumina scanning protocol

data_processing: Background substraction/ rank invariant

label: biotin

source_name: Lens

label_protocol: Standard protocol for Illumina arrays

extract_protocol: Standard total RNA isolation protocol by RNeasy mini kit (Qiagen)

value: Postnatal day 6

value: Celf1 cKO

value: Lens


title: Celf1_Lacz_P6_B

value: Control


title: 10105, hippocampus, 16p11_2_wt4


title: 10103, hippocampus, 16p11_2_wt3


title: HSC_young_LEPRpos_rep1

value: young


title: HSC_young_LEPRpos_rep2


title: X53_p7

value: Hopx-wildtype

value: type 2 multipotent progenitors (MPP2)


title: X52_p6

value: 8 weeks

value: type 3 multipotent progenitors (MPP3)


title: 10102, hippocampus, 16p11_2_wt2

value: Long Evans

value: male


title: 10110, hippocampus, 16p11_2_Del2

value: Hippocampus


title: 8-H-MN1

data_processing:
Sequenced reads were mapped to mm10 using bowtie2 (v2.3.4.3)
Gene-level expression was quantified in expected counts, FPKM and TPM using RSEM (v1.2.28)
Genome_build: mm10
Supplementary_files_format_and_content: Tab-delimited text files include expected counts, FPKM, TPM values for each sample

source_name: Hopx-/- mice BM cells + MN1

extract_protocol:
Mouse bone marrow cells were harvested from aged Hopx-wildtype and Hopx-knockout mice, and Hopx-wildtype and Hopx-knockout mice with MN1 overexpression
RNA libraries were prepared for sequencing using TruSeq Stranded mRNA Library Preparation Kit

tag: strain

value: 16 weeks

value: Hopx-knockout + MN1 overexpression


title: X120_p8

value: Hopx-knockout

value: long-term HSC (LT-HSCs)


title: Celf1_Lacz_P6_A


title: Celf1_KO_P6_B


title: 5-H-MN1

value: Bone marrow


title: MIA/luc tumor treated with PD325901 rep 4

city: Yufu

first: Naoki

last: Hijiya

state: Oita

address: 1-1, Idaigaoka Hasama-machi

department: Medicine

country: Japan

institute: Oita University

description: NOD-SCID mouse was injected with MIA/luc cells followed by treated with PD325901.  Total RNA of the tumor for expression analysis was extracted using an RNeasy Mini kit (Qiagen).

hyb_protocol: Gene Expression Hybridization kit was used for hybridization in accordance with the manufacture's instructions (Agilent Technologies).

scan_protocol: Microarray images were obtained by using a laser confocal scanner G2565BA (Agilent Technologies).

data_processing:
Feature Extraction software (Version 9.1, Agilent Technologies) with the manufacturer’s recommended settings was applied for the microarray image analysis. Analysis of the microarray images was performed with Genespring 7.3.1 Software (Agilent Technologies).
The signal intensities of probe set data were median-normalized per chip, followed by filtering based on signal intensity and flagged values.The extraction of genes upregulated or downregulated by the treatment with PD325901 was performed using the following filters: for upregulated genes, showing more that 4000 of raw data in PD325901 group and showing more than 4 times higher in PD325901 group than vehicle group, and for downregulated genes, showing more that 4000 of raw data in vehicle group and showing less than 0.25 times lower in PD325901 group than vehicle group.

source_name: Pancreatic cancer cell line

label_protocol: Cy3-labeled cRNA targets were generated using Quick Amp Labeling Kit, one-color (Agilent Technologies).

growth_protocol: MIA/luc cells were cultured in accordance with the supplier's instructions.

extract_protocol: Total RNA for expression analysis was extracted using an RNeasy Mini kit (Qiagen).

tag: agent

value: NOD-SCID


title: MIA/luc tumor treated with PD325901 rep 1


title: MIA/luc tumor treated with PD325901 rep 3


title: X119_p7


title: X118_p6


title: MIA/luc tumor treated with vehicle only rep 4

description: NOD-SCID mouse was injected with MIA/luc cells followed by treated with vehicle ony.  Total RNA of the tumor for expression analysis was extracted using an RNeasy Mini kit (Qiagen).

value: vehicle


title: 10147, hippocampus, 16p11_2_Del5

value: female


title: MIA/luc tumor treated with vehicle only rep 3


title: 4-W-MN1

source_name: Hopx WT mice BM cells + MN1

value: Hopx-wildtype + MN1 overexpression


title: MIA/luc tumor treated with PD325901 rep 2


title: X54_p8


title: 10101, hippocampus, 16p11_2_wt1

value: wt


title: 3-W-MN1


title: 10121, hippocampus, 16p11_2_Del2


title: X116_p7

value: Hopx-wildtype


title: X115_p6


title: 10145, hippocampus, 16p11_2_Del4


title: 2-W-MN1


title: 7-H-MN1


title: 10136, hippocampus, 16p11_2_Del3


title: 10120, hippocampus, 16p11_2_Del1


title: 10-H-old

source_name: Aged Hopx-/- mice BM cells

value: 18 weeks


title: X117_p8

value: 8 weeks


title: MIA/luc tumor treated with vehicle only rep 2


title: X106_p8


title: 10132, hippocampus, 16p11_2_wt5


title: 9-H-old


title: X105_p7

value: type 2 multipotent progenitors (MPP2)


title: MIA/luc tumor treated with vehicle only rep 1


title: 3-H-old


title: 10131, hippocampus, 16p11_2_wt4

value: Long Evans


title: BMSCs_hypoxia_rep2

city: Guiyang

first: Tao

last: Wang

state: Guizhou

address: 9 Beijing Road

institute: Guizhou Medical University

description:
lncRNA and mRNA
Sample cell usage per biological replicate is 50-75cm²

hyb_protocol: The labeled cRNAs were hybridized onto the Arraystar Rat LncRNA Arrays, and incubated for 17 hours at 65°C in an Agilent Hybridization Oven.

scan_protocol: After washing, slides were scanned with the Agilent Scanner G2505C.

data_processing: Data was extracted using Agilent Feature Extraction software.A series of data processing including quantile normalization were performed using the R software package. The lncRNAs that at least 7 out of 14 samples have flags in P or M (defined by GeneSpring software) were retained for further differential analyses. Differentially expressed lncRNAs with statistical significance between the two groups were identified through Volcano Plot filtering. The lncRNA/microRNA interaction was predicted with Arraystar's home-made miRNA target prediction software. All the differentially expressed lncRNAs were annotated  in detail with the lncRNA/miRNA interaction information. The lncRNA/microRNA interaction was predicted with Arraystar's home-made miRNA target prediction software. All the differentially expressed lncRNAs were annotated  in detail with the lncRNA/miRNA interaction information.

source_name: bone marrow mesenchymal stem cells

label_protocol: Total RNA from each sample was amplified and transcribed into fluorescent cRNA utilizing ramdom primer according to Arraystar Super RNA Labeling protocol (Arraystar, Inc.).

growth_protocol: BMSCs were cultured with complete L-glutamine Dulbecco’s Modified Eagle Medium (L-DMEM) containing 10% fetal bovine serum, 100 U/mL of penicillin, and 100 μg/mL of streptomycin. The cells were cultured at 37°C and 5%CO2.

extract_protocol: The culture medium was sucked out from the culture bottle and discarded, and Trizol reagent was directly added into the culture bottle. The amount of Trizol was related to the adherent area of cells, and 1-2ml Trizol was added into the adherent area of cells of about 10-15cm² (please adjust the details according to the number of cells). After repeated suction and beating for several times, the cell layer was completely dissolved visually, and the cell lysate was stored at -70℃.

treatment_protocol: The proportion of mixed gas in the cell incubator was adjusted to an oxygen concentration of 0%, nitrogen concentration of 95%, and carbon dioxide concentration of 5%. Then bone marrow mesenchymal stem cells were treated in gas mixture for 48 h to induce hypoxia, and hypoxia cell model was constructed.

value: Sprague-Dawley rat

value: bone marrow

value: hypoxia


title: 10130, hippocampus, 16p11_2_wt3

value: Hippocampus


title: X103_p7

value: Hopx-knockout


title: 10129, hippocampus, 16p11_2_wt2


title: 6-W-old

source_name: Aged Hopx WT mice BM cells


title: BMSCs_hypoxia_rep3


title: X102_p6

value: type 3 multipotent progenitors (MPP3)


title: 1-W-old

value: Bone marrow


title: BMSCs_normoxia_rep2

value: normoxia


title: 10122, hippocampus, 16p11_2_wt1


title: BMSCs_normoxia_rep3


title: X101_p8

value: long-term HSC (LT-HSCs)


title: BMSCs_normoxia_rep1


title: BMSCs_hypoxia_rep1


title: PA with 300 ug/ml nanoparticles

city: Zarqa

first: Hafez

last: Al-momani

address: Arab street

department: Microbiology, Pathology and forensic medicine

country: Jordan

institute: Hashemite university

description: Test

data_processing:
The relative expression values of QS-regulatory genes were normalized to the housekeeping gene rpoD and agarose gel electrophoresis was employed to verify the specific level of PCR amplification
For the normalization it uses the average of one housekeeping genes: rpoD
Target gene signals normalized to housekeeping genes; 2^-deltaCt, where deltaCt = (Ct_Target − Ct_HKG)]
we performs all deltadeltaCt based fold-change calculations from the uploaded raw threshold cycle data.
Fold Change worksheet reports test/control (i.e., with nanoparticles/PA without nanopartices) ratios.

label: SYBR Green

organism: Pseudomonas aeruginosa

source_name: Bacteria

label_protocol: Quantitative polymerase chain reaction (qPCR) was conducted with a BIO-RAD Thermal cycler; for amplification, 2 𝜇L of template DNA was combined with 0.5 𝜇L of each forward and reverse primer, 10 𝜇L of Luna Universal qPCR Master Mix, and 7 𝜇L of nuclease-free water. The resultant reaction volume was 20 𝜇L.

extract_protocol: An RNeasy Mini Kit (Qiagen, , Germany) was used to extract the biofilm RNA

tag: activation agent

value: treated

tag: time

value: 24 hours


title: PA without nanoparticles

description: Control

value: control


title: X104_p8


title: PA with 900 ug/ml nanoparticles

description: Test


title: 7-W-old

value: Hopx-wildtype


title: GBM EC, 5465-sgPHGDH

status: Public on Feb 17 2023

city: Philadelphia

first: Yi

last: Fan

state: PA

address: 3400 Civic Center Blvd, SCTR 8-168

department: Radiation Oncology

institute: University of Pennsylvania

data_processing:
Kallisto pseudo-alignment quantification was performed for the RNA-Seq data using Kallisto version 0.46.1 obtained from github. Kallisto index was built with reference transcriptome GRCh38.
In R (version 4.0.5), transcripts were collapsed to genes using package tximport (version 1.22.0), and then normalized and filtered by edgeR (version 3.36.0) to generate a count matrix.
Assembly: GRCh38
Supplementary files format and content: Raw count matrix in a comma-separated values file

source_name: Human tumor (GBM) EC

growth_protocol: GBM-derived ECs were maintained in Endothelial Cell Medium (ECM, ScienCell, supplemented with VEGF-A) at 37 degrees Celcius with 5% CO2. All cells were used between passages two and five.

extract_protocol:
2 x 10^6 cells were lysed in 1 ml TRIzol (Thermo Fisher), followed by RNA extraction. The isolated RNA was purified with an RNeasy Plus Mini Kit (Qiagen).
The DNA library was constructed by using a RiboZero protocol with TapeStation (Agilent), and subjected to deep sequencing (125 PE, approximately 40 megabyte reads for each sample, Illumina HiSeq 2500) at the Children’s Hospital of Philadelphia.

treatment_protocol: The gRNA sequence (5’-CACCGGAGGCCGACAATGACTGCG-3’) targeting PHGDH or control gRNA sequence was subcloned into lentiCRISPRv2 puro vector. ECs were transduced with the lenvirus expressing PHGDH sgRNA, followed by cell selection by incubation with medium containing 2 µg/ml puromycin.

value: glioblastoma (GBM)

value: GBM tumor

value: GBM endothelial cell

value: sgPHGDH

value: PHGDH knockout


title: GBM EC, 5391-sgPHGDH


title: GBM EC, 5377-sgPHGDH


title: GBM EC, 5465-sgControl

value: sgControl


title: GBMEC5785

description: Primary tumor vascular endothelial from patient #5785

source_name: Patient #5785

extract_protocol:
2 x 10^6 ECs were lysed in 1 ml TRIzol (Thermo Fisher Scientific), followed by RNA extraction according to the manufacturer’s instructions. The isolated RNA was purified with RNeasy Plus Mini Kit (Qiagen).
After a quality control step using RNA Nano assay chips with a 2100 Bioanalyzer (Agilent), a library was constructed using a Ribo-Zero protocol with TapeStation (Agilent) and subjected to deep sequencing (125 paired-end (PE); approximately 40 MB reads for each sample; Illumina HiSeq 2500 sequencer) at the Children’s Hospital of Philadelphia.

value: Primary tumor vascular endothelial cell


title: GBMEC5441

description: Primary tumor vascular endothelial from patient #5441

source_name: Patient #5441


title: GBMEC5433

description: Primary tumor vascular endothelial from patient #5433

source_name: Patient #5433


title: GBMEC5391

description: Primary tumor vascular endothelial from patient #5391

source_name: Patient #5391


title: 180 rDNA strain, 45 min S phase

type: genomic

city: Seattle

first: Mosur

last: Raghuraman

address: Foege Bldg, 3720 15th Ave NE

department: Genome Sciences Box 355065

institute: University of Washington

description: HH and HL DNA were separated by CsCl density gradient centrifugation

hyb_protocol: Hybridization and washing were done using standard conditions specified by Agilent (using Agilent Blocking Agent and HI-rPM hybridization buffer).

scan_protocol: Arrays were scanned on an Agilent Technologies Scanner G2505C and features were extracted using protocol CGH 105 Dec08 CP in feature extractor (FE) v. 10.7.3.1

data_processing: After extraction of hybridization intensities, we used the known % replication values from the slot blot analysis to normalize the overall % replication in each sample, with an added correction for the percentage of cells in the population that successfully completed S phase (obtained from the maximum % replication seen at the end of S phase from the slot blot analysis).

organism: Saccharomyces cerevisiae

source_name: 45 min S phase, HH (unreplicated) DNA, 180 copies rDNA

label_protocol: 200 ng of HH or HL DNA were labeled with Cy3 and Cy5, respectively, using random-primed synthesis with exonuclease-deficient Klenow.

growth_protocol: We grew cells in isotopically dense (13C-glucose, 15N-ammonium sulfate) for ≥7 population doublings.  We then arrested the cells in G1 using a-factor, filtered and resuspended the cells in isotopically light (12C, 14N) medium.

extract_protocol: DNA was extracted using glass beads and phenol/chloroform (standard smash-and-grab method).  The DNA was digested with EcoRI.  Heavy-heavy (HH) and heavy-light (HL) DNA were separated by ultracentrifugation in cesium chloride gradients.  The gradients were drip-fractionated.  HH and HL portions of the gradient were identified by slot blot hybridization.  The DNA was recovered from CsCl by ethanol precipitation.

treatment_protocol: Cells were released into S phase by addition of Pronase.  We collected samples at intervals through the ensuing S phase (30,35,40,45,50 min for 35 rDNA strain; 25,30,35,40,45,55 min for 180 rDNA strain).

tag: cell cycle phase

value: S phase

tag: s phase time

value: 45 min

tag: fraction

value: HH (unreplicated) DNA

source_name: 45 min S phase, HL (replicated) DNA, 180 copies rDNA

value: HL (replicated) DNA


title: GBMEC5377

description: Primary tumor vascular endothelial from patient #5377

source_name: Patient #5377

tag: tissue


title: 35 rDNA strain, 50 min S phase

source_name: 50 min S phase, HH (unreplicated) DNA, 35 copies rDNA

value: 50 min

source_name: 50 min S phase, HL (replicated) DNA, 35 copies rDNA


title: Normal EC-2

description: Brain vascular endothelial cell line

source_name: Normal human brain

value: normal

value: brain

tag: cell type

value: Brain vascular endothelial cell


title: Normal EC-3

description: Brain vascular endothelial cell line


title: 180 rDNA strain, 40 min S phase

source_name: 40 min S phase, HH (unreplicated) DNA, 180 copies rDNA

value: 40 min

source_name: 40 min S phase, HL (replicated) DNA, 180 copies rDNA


title: 180 rDNA strain, 30 min S phase

source_name: 30 min S phase, HH (unreplicated) DNA, 180 copies rDNA

value: 30 min

source_name: 30 min S phase, HL (replicated) DNA, 180 copies rDNA


title: NINT Female rep2

city: Jeonju

first: Mousumee

last: Khan

address: 20 Geonji-ro

country: South Korea

institute: Jeonbuk National University

data_processing:
The read count data were processed based on geometric median and ratio normalization method using DESeq2 package within R program
All the read count was imported into DESeq2 and read counts were log transformed to validate the data quality by the hierarchical clustering method
All normalized counts were used for further analysis. DEG’s were identified by Wald test in DESeq2 package by using three pair of group comparison: 1) INT-F0, 2) NINT-F0, 3) INT-NINT, filter criterion was an adjusted P value <0.01
To better understand the functional involvements of the DEGs, the latest Ensembl release annotation set of D. melanogaster (AH89395) was used for gene level annotation in AnnotationHub package
Gene ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) associated with the significant genes were performed by the clusterProfiler package with cut-off values as adjusted P value < 0.05.  visualization of Gene Set Enrichment Analysis (GSEA).
The enrichplot package  was used for visualization of Gene Set Enrichment Analysis (GSEA)
Assembly: dm6
Supplementary files format and content: Matrix table with mRNA-seq Read Count of every gene of all male samples
Supplementary files format and content: Matrix table with mRNA-seq Read Count of every gene of all female samples

organism: Drosophila melanogaster

source_name: Whole fly

extract_protocol:
total RNA was isolated using Trizol reagent (Invitrogen) according to the manufacturer's protocol. Libraries were prepared from total RNA using the SMARTer Stranded RNA-Seq Kit (Clontech Laboratories, Inc., Cat# 634837)
mRNA libraries were prepared for sequencing using standard Illumina protocols

value: genetically heterogenous

value: Whole fly

value: 5 days

tag: group

value: NINT

value: Female


title: NINT Female rep3


title: CD90+HSP91+ HCC cell

city: beijing

first: Xiong

last: Shu

address: No.31, Xinjiekou East Street, Xicheng District, Beijing

institute: Beijing Research Institute of Traumatology and Orthopaedics, Beijing JiShuiTan Hospital

description: Gene expression data from normal CD90+HSP90- HCC cell

hyb_protocol: Samples were hybridized using Affymetrix hybridization kit materials. Heat cocktails at 99°C for 5 minutes, then 42°C for 5 minutes ,then centrifuge at 13200 rpm for 1 minute. Transfer 200μl of hyb solution to each array, then tape holes and parafilm. Hybridize 16 hours at 45°C at 60rpm. Fluidics washing in Fluidics Station 450.

scan_protocol: GeneChips were scanned using the Affymetrix Gene ChIp Scanner 3000.

data_processing: The data were analyzed with Microarray Suite version 3.0 (MAS 3.0).

source_name: normal CD90+HSP91+ HCC cell

growth_protocol: CD90+HSP90+ HCC cells

extract_protocol: Trizol extraction of total RNA was performed according to the manufacturer's instructions.In vitro amplification and biotin labeling with Ambion # 1792 cRNA amplification kit

treatment_protocol: Cultured sphere cells, sphere cells were harvested and dissociated into single cells with trypsin for the subsequent experiments.


title: CD90+HSP90+ HCC cell, 11C9 treated

description: Gene expression data from 11C9_treated CD90+HSP90+ HCC cell

source_name: 11C9_treated CD90+HSP90+ HCC cell

label_protocol: In vitro amplification and biotin labeling with Ambion # 1792 cRNA amplification kit


title: 180 rDNA strain, 55 min S phase

source_name: 55 min S phase, HH (unreplicated) DNA, 180 copies rDNA

value: 55 min

source_name: 55 min S phase, HL (replicated) DNA, 180 copies rDNA


title: 35 rDNA strain, 40 min S phase

source_name: 40 min S phase, HH (unreplicated) DNA, 35 copies rDNA

source_name: 40 min S phase, HL (replicated) DNA, 35 copies rDNA


title: 180 rDNA strain, 35 min S phase

source_name: 35 min S phase, HH (unreplicated) DNA, 180 copies rDNA

value: 35 min

source_name: 35 min S phase, HL (replicated) DNA, 180 copies rDNA


title: 35 rDNA strain, 30 min S phase

source_name: 30 min S phase, HH (unreplicated) DNA, 35 copies rDNA

value: S phase

source_name: 30 min S phase, HL (replicated) DNA, 35 copies rDNA


title: 35 rDNA strain, 35 min S phase

source_name: 35 min S phase, HH (unreplicated) DNA, 35 copies rDNA

source_name: 35 min S phase, HL (replicated) DNA, 35 copies rDNA


title: 35 rDNA strain, 45 min S phase

source_name: 45 min S phase, HH (unreplicated) DNA, 35 copies rDNA

value: 45 min

source_name: 45 min S phase, HL (replicated) DNA, 35 copies rDNA


title: NINT Female rep1


title: 180 rDNA strain, 25 min S phase

source_name: 25 min S phase, HH (unreplicated) DNA, 180 copies rDNA

value: 25 min

value: HH (unreplicated) DNA

source_name: 25 min S phase, HL (replicated) DNA, 180 copies rDNA


title: NINT Male rep3

source_name: Whole fly

value: Male


title: GBM EC, 5391-sgControl

source_name: Human tumor (GBM) EC

value: glioblastoma (GBM)

value: GBM tumor

value: GBM endothelial cell

tag: treatment

value: sgControl

tag: genotype

value: control


title: GBM EC, 5377-sgControl

tag: disease state


title: INT Female rep3

value: genetically heterogenous

tag: age

value: INT


title: NINT Male rep1

value: Whole fly

value: 5 days


title: NINT Male rep2

value: NINT


title: INT Female rep2

value: Female


title: INT Female rep1


title: INT Male rep2


title: Normal EC-1

description: Brain vascular endothelial cell line

source_name: Normal human brain

value: normal

value: brain

value: Brain vascular endothelial cell


title: INT Male rep1


title: F0 Female rep3

value: Oregon-R-C, DGRP-100, BER_2, Harwich, Canton-S, Wild_1A, w1118, multiple wing hairs (mwh), flare3 (flr3), Oregon R flare3 (ORR, flr3)


title: F0 Male rep3


title: F0 Male rep1


title: F0 Female rep1


title: F0 Male rep2


title: F0 Female rep2


title: INT Male rep3


title: NCC-HN220M [RHT5076]

city: Singapore

first: Yiqun Elaine

last: Cao

address: 8 College Road

country: Singapore

institute: Duke-NUS Medical School

description:
C1 Fluidigm
good_counts.RData
good_tpm.RData
good_cellinfo.RData

data_processing:
The raw reads in FASTQ files were aligned to the human genome (hg19 assembly) using STAR v2.6.0
Quantification of gene expression using RSEM v1.3.0 to generate TPM values
Cells with (i) less than 100,000 raw counts, (ii) less than 10% unique exonic mapping rate, or (iii) less than 1,000 expressed genes (TPM > 1) were excluded from analysis
Assembly: hg19
Supplementary files format and content: good_counts.RData is a R data file that contains a data frame of raw read counts for 1,317 cells passing the quality control
Supplementary files format and content: good_tpm.RData is a R data file that contains a data frame of TPM gene expression values for 1,317 cells passing the quality control
Supplementary files format and content: good_cellinfo.RData is a R data file that contains a data frame of cell information for 1,317 cells passing the quality control

source_name: tumor

extract_protocol:
Tumors were removed, processed into single cell suspension using a GentleMACS dissociator (Miltenyi Biotec) as described in the manufacturer protocol. Cell suspension were plated and passaged until a tumor majority was observed. On the day of experiment, cancer cell culture were trypsinized and single cell was captured, lysed and reverse transcribed into cDNA using Fluidigm Integrated Fluidic Circuit (IFC) and a Fluidigm C1 instrument (Fluidigm, South San Francisco, CA).
Libraries were prepared for sequencing using C1 Single-Cell Reagent Kit for mRNA Seq

value: tumor

tag: patient id

tag: primary site

value: L Lateral Tongue

tag: lymph node_involved

value: Yes

tag: Stage

tag: grade

tag: site of_origin

value: Metastatic (nodal)

tag: cell line

value: patient-derived cell culture

value: cancer cell


title: NCC-HN220M [RHT5072]


title: NCC-HN220M [RHT5081]


title: NCC-HN220M [RHT5075]


title: NCC-HN220M [RHT5084]


title: NCC-HN220M [RHT5074]


title: NCC-HN220M [RHT5079]


title: NCC-HN220M [RHT5073]


title: NCC-HN220M [RHT5071]


title: NCC-HN220M [RHT5085]


title: NCC-HN220M [RHT5082]


title: NCC-HN220M [RHT5070]


title: NCC-HN220M [RHT5077]


title: NCC-HN220M [RHT5083]


title: NCC-HN220M [RHT5069]


title: NCC-HN220M [RHT5078]


title: NCC-HN220M [RHT5086]


title: NCC-HN220M [RHT5066]


title: NCC-HN220M [RHT5068]


title: NCC-HN220M [RHT5065]


title: NCC-HN220M [RHT5080]


title: NCC-HN220M [RHT5064]


title: NCC-HN220M [RHT5063]


title: NCC-HN220M [RHT5062]


title: NCC-HN220M [RHT5059]


title: NCC-HN220M [RHT5060]


title: NCC-HN220M [RHT5061]


title: NCC-HN220M [RHT5067]


title: NCC-HN220M [RHT5057]


title: NCC-HN220M [RHT5041]


title: NCC-HN220M [RHT5048]


title: NCC-HN220M [RHT5049]


title: NCC-HN220M [RHT5044]


title: NCC-HN220M [RHT5052]


title: NCC-HN220M [RHT5054]


title: NCC-HN220M [RHT5038]


title: NCC-HN220M [RHT5056]


title: NCC-HN220M [RHT5055]


title: NCC-HN220M [RHT5050]


title: NCC-HN220M [RHT5045]


title: NCC-HN220M [RHT5046]


title: NCC-HN220M [RHT5053]


title: NCC-HN220M [RHT5040]


title: NCC-HN220M [RHT5037]


title: NCC-HN220M [RHT5047]


title: NCC-HN220M [RHT5039]


title: NCC-HN220M [RHT5035]


title: NCC-HN220M [RHT5058]


title: NCC-HN220M [RHT5032]


title: NCC-HN220M [RHT5033]


title: NCC-HN220M [RHT5051]


title: NCC-HN220M [RHT5030]


title: NCC-HN220M [RHT5031]


title: NCC-HN220M [RHT5036]


title: NCC-HN220M [RHT5034]


title: NCC-HN220M [RHT5029]


title: NCC-HN220M [RHT5042]


title: NCC-HN220M [RHT5043]


title: NCC-HN220M [RHT5028]


title: NCC-HN220M [RHT5027]


title: NCC-HN220M [RHT5026]


title: NCC-HN220M [RHT5025]


title: NCC-HN220M [RHT5023]


title: NCC-HN220P [RHT5019]

value: Primary


title: NCC-HN220P [RHT5020]


title: NCC-HN220P [RHT5016]


title: NCC-HN220P [RHT5013]


title: NCC-HN220P [RHT5012]


title: NCC-HN220P [RHT5015]


title: NCC-HN220P [RHT5007]


title: NCC-HN220P [RHT5021]


title: NCC-HN220P [RHT5011]


title: NCC-HN220P [RHT5017]


title: NCC-HN220P [RHT5004]


title: NCC-HN220P [RHT5008]


title: NCC-HN220P [RHT5006]


title: NCC-HN220P [RHT5002]


title: NCC-HN220P [RHT5003]


title: NCC-HN220P [RHT5009]


title: NCC-HN220P [RHT5014]


title: NCC-HN220P [RHT5005]


title: NCC-HN220M [RHT5024]


title: NCC-HN220P [RHT5001]


title: NCC-HN220P [RHT4998]


title: NCC-HN220P [RHT5000]


title: NCC-HN220P [RHT5022]


title: NCC-HN220P [RHT4999]


title: NCC-HN220P [RHT5018]


title: NCC-HN220P [RHT4997]


title: NCC-HN220P [RHT5010]


title: NCC-HN220P [RHT4995]


title: NCC-HN220P [RHT4994]


title: NCC-HN220P [RHT4992]


title: NCC-HN220P [RHT4993]


title: NCC-HN220P [RHT4989]


title: NCC-HN220P [RHT4985]


title: NCC-HN220P [RHT4986]


title: NCC-HN220P [RHT4990]


title: NCC-HN220P [RHT4987]


title: NCC-HN220P [RHT4980]


title: NCC-HN220P [RHT4983]


title: NCC-HN220P [RHT4981]


title: NCC-HN220P [RHT4982]


title: NCC-HN220P [RHT4978]


title: NCC-HN220P [RHT4979]


title: NCC-HN220P [RHT4976]


title: NCC-HN220P [RHT4977]


title: NCC-HN220P [RHT4988]


title: NCC-HN220P [RHT4984]


title: NCC-HN220P [RHT4972]


title: NCC-HN220P [RHT4970]


title: NCC-HN220P [RHT4973]


title: NCC-HN220P [RHT4974]


title: NCC-HN220P [RHT4971]


title: NCC-HN220P [RHO1580]


title: NCC-HN220P [RHT4975]


title: NCC-HN220P [RHO1579]


title: NCC-HN220P [RHO1578]


title: NCC-HN220P [RHT4991]


title: NCC-HN220P [RHO1577]


title: NCC-HN220P [RHT4996]


title: NCC-HN220P [RHO1575]


title: NCC-HN220P [RHO1576]


title: NCC-HN220P [RHO1574]


title: NCC-HN220P [RHO1573]


title: NCC-HN220P [RHO1572]


title: NCC-HN220P [RHO1571]


title: NCC-HN220P [RHO1570]


title: NCC-HN220P [RHO1569]


title: NCC-HN220P [RHO1562]


title: NCC-HN220P [RHO1568]


title: NCC-HN220P [RHO1564]


title: NCC-HN220P [RHO1567]


title: NCC-HN220P [RHO1566]


title: NCC-HN220P [RHO1563]


title: NCC-HN220P [RHO1558]


title: NCC-HN220P [RHO1557]


title: NCC-HN220P [RHO1556]


title: NCC-HN220P [RHO1565]


title: NCC-HN220P [RHO1554]


title: NCC-HN220P [RHO1553]


title: NCC-HN220P [RHO1555]


title: NCC-HN220P [RHO1560]


title: NCC-HN220P [RHO1559]


title: NCC-HN220P [RHO1552]


title: NCC-HN217M [RHO1482]

value: R Tongue

value: Yes Level 1 & Level 2


title: NCC-HN217M [RHO1483]


title: NCC-HN217M [RHO1484]


title: NCC-HN220P [RHO1561]


title: NCC-HN217M [RHO1480]


title: NCC-HN217M [RHO1481]


title: NCC-HN217M [RHO1478]


title: NCC-HN217M [RHO1477]


title: NCC-HN217M [RHO1476]


title: NCC-HN217M [RHO1475]


title: NCC-HN217M [RHO1474]


title: NCC-HN217M [RHO1473]


title: NCC-HN220P [RHO1551]


title: NCC-HN217M [RHO1464]


title: NCC-HN217M [RHO1466]


title: NCC-HN217M [RHO1465]


title: NCC-HN217M [RHO1468]


title: NCC-HN217M [RHO1459]


title: NCC-HN217M [RHO1462]


title: NCC-HN217M [RHO1469]


title: NCC-HN217M [RHO1470]


title: NCC-HN217M [RHO1461]


title: NCC-HN217M [RHO1479]


title: NCC-HN217M [RHO1460]


title: NCC-HN217M [RHO1457]


title: NCC-HN217M [RHO1455]


title: NCC-HN217M [RHO1456]


title: NCC-HN217M [RHO1451]


title: NCC-HN217M [RHO1452]


title: NCC-HN217M [RHO1454]


title: NCC-HN217M [RHO1453]


title: NCC-HN217M [RHO1449]


title: NCC-HN217M [RHO1448]


title: NCC-HN217M [RHO1450]


title: NCC-HN217M [RHO1471]


title: NCC-HN217M [RHO1472]


title: NCC-HN217M [RHO1445]


title: NCC-HN217M [RHO1467]


title: NCC-HN217M [RHO1446]


title: NCC-HN217M [RHO1463]


title: NCC-HN217M [RHO1444]


title: NCC-HN217M [RHO1441]


title: NCC-HN217M [RHO1443]


title: NCC-HN217M [RHO1458]


title: NCC-HN217M [RHO1442]


title: NCC-HN217M [RHO1436]


title: NCC-HN217M [RHO1438]


title: NCC-HN217M [RHO1437]


title: NCC-HN217M [RHO1439]


title: NCC-HN217M [RHO1440]


title: NCC-HN217M [RHO1435]


title: NCC-HN217M [RHO1432]


title: NCC-HN217M [RHO1430]


title: NCC-HN217M [RHO1434]


title: NCC-HN217M [RHO1429]


title: NCC-HN217M [RHO1427]


title: NCC-HN217M [RHO1428]


title: NCC-HN217M [RHO1431]


title: NCC-HN217M [RHO1426]


title: NCC-HN217M [RHO1447]


title: NCC-HN217M [RHO1433]


title: NCC-HN217M [RHO1425]


title: NCC-HN217M [RHO1423]


title: NCC-HN217M [RHO1422]


title: NCC-HN217M [RHO1421]


title: NCC-HN217M [RHO1419]


title: NCC-HN217M [RHO1420]


title: NCC-HN217M [RHO1418]


title: NCC-HN217M [RHO1417]


title: NCC-HN217M [RHO1416]


title: NCC-HN217M [RHO1414]


title: NCC-HN217M [RHO1415]


title: NCC-HN217M [RHO1413]


title: NCC-HN217M [RHO1412]


title: NCC-HN217M [RHO1410]


title: NCC-HN217M [RHO1411]


title: NCC-HN217M [RHO1424]


title: NCC-HN217M [RHO1408]


title: NCC-HN217M [RHO1404]


title: NCC-HN217M [RHO1402]


title: NCC-HN217M [RHO1396]


title: NCC-HN217M [RHO1399]


title: NCC-HN217M [RHO1395]


title: NCC-HN217M [RHO1405]


title: NCC-HN217M [RHO1403]


title: NCC-HN217M [RHO1406]


title: NCC-HN217M [RHO1400]


title: NCC-HN217M [RHO1397]


title: NCC-HN217M [RHO1401]


title: NCC-HN217M [RHO1393]


title: NCC-HN217M [RHO1392]


title: NCC-HN217M [RHO1394]


title: NCC-HN217M [RHO1391]


title: NCC-HN217M [RHO1389]


title: NCC-HN217P [RHO1550]


title: NCC-HN217P [RHO1549]


title: NCC-HN217M [RHO1409]


title: NCC-HN217P [RHO1548]


title: NCC-HN217P [RHO1547]


title: NCC-HN217M [RHO1407]


title: NCC-HN217P [RHO1545]


title: NCC-HN217P [RHO1546]


title: NCC-HN217P [RHO1544]


title: NCC-HN217M [RHO1398]


title: NCC-HN217P [RHO1543]


title: NCC-HN217P [RHO1542]


title: NCC-HN217P [RHO1538]


title: NCC-HN217P [RHO1540]


title: NCC-HN217M [RHO1390]


title: NCC-HN217P [RHO1537]


title: NCC-HN217P [RHO1539]


title: NCC-HN217P [RHO1535]


title: NCC-HN217P [RHO1541]


title: NCC-HN217P [RHO1529]


title: NCC-HN217P [RHO1533]


title: NCC-HN217P [RHO1530]


title: NCC-HN217P [RHO1534]


title: NCC-HN217P [RHO1531]


title: NCC-HN217P [RHO1526]


title: NCC-HN217P [RHO1524]


title: NCC-HN217P [RHO1525]


title: NCC-HN217P [RHO1528]


title: NCC-HN217P [RHO1527]


title: NCC-HN217P [RHO1520]


title: NCC-HN217P [RHO1519]


title: NCC-HN217P [RHO1517]


title: NCC-HN217P [RHO1521]


title: NCC-HN217P [RHO1523]


title: NCC-HN217P [RHO1514]


title: NCC-HN217P [RHO1518]


title: NCC-HN217P [RHO1515]


title: NCC-HN217P [RHO1536]


title: NCC-HN217P [RHO1516]


title: NCC-HN217P [RHO1513]


title: NCC-HN217P [RHO1532]


title: NCC-HN217P [RHO1522]


title: NCC-HN217P [RHO1512]


title: NCC-HN217P [RHO1509]


title: NCC-HN217P [RHO1508]


title: NCC-HN217P [RHO1510]


title: NCC-HN217P [RHO1504]


title: NCC-HN217P [RHO1506]


title: NCC-HN217P [RHO1505]


title: NCC-HN217P [RHO1507]


title: NCC-HN217P [RHO1501]


title: NCC-HN217P [RHO1503]


title: NCC-HN217P [RHO1502]


title: NCC-HN217P [RHO1497]


title: NCC-HN217P [RHO1500]


title: NCC-HN217P [RHO1499]


title: NCC-HN217P [RHO1494]


title: NCC-HN217P [RHO1495]


title: NCC-HN217P [RHO1491]


title: NCC-HN217P [RHO1489]


title: NCC-HN217P [RHO1487]


title: NCC-HN217P [RHO1488]


title: NCC-HN217P [RHO1490]


title: NCC-HN217P [RHO1493]


title: NCC-HN217P [RHO1486]


title: NCC-HN160M [RHH7439]

value: R Dorsal tongue,  posterior-lateral


title: NCC-HN160M [RHH7437]


title: NCC-HN160M [RHH7438]


title: NCC-HN217P [RHO1485]


title: NCC-HN217P [RHO1511]


title: NCC-HN160M [RHH7435]


title: NCC-HN160M [RHH7436]


title: NCC-HN217P [RHO1498]


title: NCC-HN160M [RHH7434]


title: NCC-HN160M [RHH7433]


title: NCC-HN217P [RHO1496]


title: NCC-HN217P [RHO1492]


title: NCC-HN160M [RHH7424]


title: NCC-HN160M [RHH7431]


title: NCC-HN160M [RHH7430]


title: NCC-HN160M [RHH7427]


title: NCC-HN160M [RHH7425]


title: NCC-HN160M [RHH7423]


title: NCC-HN160M [RHH7429]


title: NCC-HN160M [RHH7426]


title: NCC-HN160M [RHH7428]


title: NCC-HN160M [RHH7422]


title: NCC-HN160M [RHH7421]


title: NCC-HN160M [RHH7418]


title: NCC-HN160M [RHH7419]


title: NCC-HN160M [RHH7416]


title: NCC-HN160M [RHH7420]


title: NCC-HN160M [RHH7417]


title: NCC-HN160M [RHH7414]


title: NCC-HN160M [RHH7412]


title: NCC-HN160M [RHH7408]


title: NCC-HN160M [RHH7415]


title: NCC-HN160M [RHH7432]


title: NCC-HN160M [RHH7411]


title: NCC-HN160M [RHH7409]


title: NCC-HN160M [RHH7410]


title: NCC-HN160M [RHH7406]


title: NCC-HN160M [RHH7405]


title: NCC-HN160M [RHH7407]


title: NCC-HN160M [RHH7404]


title: NCC-HN160M [RHH7401]


title: NCC-HN160M [RHH7400]


title: NCC-HN160M [RHH7399]


title: NCC-HN160M [RHH7397]


title: NCC-HN160M [RHH7398]


title: NCC-HN160M [RHH7402]


title: NCC-HN160M [RHH7413]


title: NCC-HN160M [RHH7392]


title: NCC-HN160M [RHH7403]


title: NCC-HN160M [RHH7391]


title: NCC-HN160M [RHH7394]


title: NCC-HN160M [RHH7396]


title: NCC-HN160M [RHH7389]


title: NCC-HN160M [RHH7395]


title: NCC-HN160M [RHH7386]


title: NCC-HN160M [RHH7384]


title: NCC-HN160M [RHH7390]


title: NCC-HN160M [RHH7385]


title: NCC-HN160M [RHH7383]


title: NCC-HN160M [RHH7381]


title: NCC-HN160M [RHH7380]


title: NCC-HN160M [RHH7379]


title: NCC-HN160M [RHH7382]


title: NCC-HN160M [RHH7377]


title: NCC-HN160M [RHH7378]


title: NCC-HN160M [RHH7376]


title: NCC-HN160M [RHH7393]


title: NCC-HN160M [RHH7375]


title: NCC-HN160M [RHH7374]


title: NCC-HN160M [RHH7371]


title: NCC-HN160M [RHH7372]


title: NCC-HN160M [RHH7369]


title: NCC-HN160M [RHH7388]


title: NCC-HN160M [RHH7370]


title: NCC-HN160M [RHH7387]


title: NCC-HN160M [RHH7367]


title: NCC-HN160M [RHH7368]


title: NCC-HN160M [RHH7366]


title: NCC-HN160M [RHH7365]


title: NCC-HN160M [RHH7362]


title: NCC-HN160M [RHH7363]


title: NCC-HN160M [RHH7359]


title: NCC-HN160M [RHH7364]


title: NCC-HN160M [RHH7358]


title: NCC-HN160M [RHH7361]


title: NCC-HN160M [RHH7356]


title: NCC-HN160M [RHH7357]


title: NCC-HN160M [RHH7360]


title: NCC-HN160M [RHH7351]


title: NCC-HN160M [RHH7353]


title: NCC-HN160M [RHH7354]


title: NCC-HN160M [RHH7352]


title: NCC-HN160M [RHH7349]


title: NCC-HN160M [RHH7348]


title: NCC-HN160M [RHH7347]


title: NCC-HN160M [RHH7346]


title: NCC-HN160M [RHH7345]


title: NCC-HN160M [RHH7344]


title: NCC-HN160P [RHH7341]


title: NCC-HN160P [RHH7342]


title: NCC-HN160P [RHH7339]


title: NCC-HN160P [RHH7340]


title: NCC-HN160P [RHH7338]


title: NCC-HN160P [RHH7336]


title: NCC-HN160P [RHH7335]


title: NCC-HN160P [RHH7337]


title: NCC-HN160P [RHH7334]


title: NCC-HN160P [RHH7332]


title: NCC-HN160M [RHH7355]


title: NCC-HN160P [RHH7331]


title: NCC-HN160P [RHH7329]


title: NCC-HN160P [RHH7327]


title: NCC-HN160P [RHH7326]


title: NCC-HN160P [RHH7328]


title: NCC-HN160P [RHH7325]


title: NCC-HN160M [RHH7350]


title: NCC-HN160P [RHH7323]


title: NCC-HN160P [RHH7322]


title: NCC-HN160P [RHH7343]


title: NCC-HN160P [RHH7318]


title: NCC-HN160P [RHH7317]


title: NCC-HN160P [RHH7321]


title: NCC-HN160P [RHH7319]


title: NCC-HN160P [RHH7320]


title: NCC-HN160P [RHH7315]


title: NCC-HN160P [RHH7333]


title: NCC-HN160P [RHH7316]


title: NCC-HN160P [RHH7330]


title: NCC-HN160P [RHH7313]


title: NCC-HN160P [RHH7314]


title: NCC-HN160P [RHH7324]


title: NCC-HN160P [RHH7299]


title: NCC-HN160P [RHH7305]


title: NCC-HN160P [RHH7303]


title: NCC-HN160P [RHH7312]


title: NCC-HN160P [RHH7307]


title: NCC-HN160P [RHH7296]


title: NCC-HN160P [RHH7311]


title: NCC-HN160P [RHH7302]


title: NCC-HN160P [RHH7295]


title: NCC-HN160P [RHH7297]


title: NCC-HN160P [RHH7300]


title: NCC-HN160P [RHH7294]


title: NCC-HN160P [RHH7310]


title: NCC-HN160P [RHH7304]


title: NCC-HN160P [RHH7298]


title: NCC-HN160P [RHH7301]


title: NCC-HN160P [RHH7306]


title: NCC-HN160P [RHH7293]


title: NCC-HN160P [RHH7291]


title: NCC-HN160P [RHH7292]


title: NCC-HN160P [RHH7290]


title: NCC-HN160P [RHH7289]


title: NCC-HN160P [RHH7287]


title: NCC-HN160P [RHH7286]


title: NCC-HN160P [RHH7285]


title: NCC-HN160P [RHH7284]


title: NCC-HN160P [RHH7282]


title: NCC-HN160P [RHH7281]


title: NCC-HN160P [RHH7283]


title: NCC-HN160P [RHH7280]


title: NCC-HN160P [RHH7278]


title: NCC-HN160P [RHH7279]


title: NCC-HN160P [RHH7276]


title: NCC-HN160P [RHH7275]


title: NCC-HN160P [RHH7277]


title: NCC-HN160P [RHH7266]


title: NCC-HN160P [RHH7265]


title: NCC-HN160P [RHH7267]


title: NCC-HN160P [RHH7272]


title: NCC-HN160P [RHH7274]


title: NCC-HN160P [RHH7273]


title: NCC-HN160P [RHH7271]


title: NCC-HN160P [RHH7264]


title: NCC-HN160P [RHH7270]


title: NCC-HN160P [RHH7269]


title: NCC-HN160P [RHH7263]


title: NCC-HN160P [RHH7259]


title: NCC-HN160P [RHH7260]


title: NCC-HN160P [RHH7261]


title: NCC-HN160P [RHH7258]


title: NCC-HN160P [RHH7257]


title: NCC-HN160P [RHH7255]


title: NCC-HN160P [RHH7253]


title: NCC-HN160P [RHH7252]


title: NCC-HN160P [RHH7268]


title: NCC-HN160P [RHH7250]


title: NCC-HN160P [RHH7262]


title: NCC-HN160P [RHH7256]


title: NCC-HN159M [RHH7243]


title: NCC-HN159M [RHH7247]


title: NCC-HN159M [RHH7246]


title: NCC-HN160P [RHH7249]


title: NCC-HN160P [RHH7248]


title: NCC-HN160P [RHH7254]


title: NCC-HN159M [RHH7245]


title: NCC-HN159M [RHH7230]


title: NCC-HN159M [RHH7236]


title: NCC-HN159M [RHH7239]


title: NCC-HN159M [RHH7240]


title: NCC-HN159M [RHH7232]


title: NCC-HN159M [RHH7237]


title: NCC-HN159M [RHH7235]


title: NCC-HN159M [RHH7231]


title: NCC-HN159M [RHH7234]


title: NCC-HN159M [RHH7241]


title: NCC-HN159M [RHH7242]


title: NCC-HN160P [RHH7251]


title: NCC-HN159M [RHH7229]


title: NCC-HN159M [RHH7233]


title: NCC-HN160P [RHH7288]


title: NCC-HN159M [RHH7228]


title: NCC-HN159M [RHH7244]


title: NCC-HN159M [RHH7238]


title: NCC-HN159M [RHH7226]


title: NCC-HN159M [RHH7225]


title: NCC-HN159M [RHH7227]


title: NCC-HN159M [RHH7224]


title: NCC-HN159M [RHH7223]


title: NCC-HN159M [RHH7222]


title: NCC-HN159M [RHH7220]


title: NCC-HN159M [RHH7218]


title: NCC-HN159M [RHH7221]


title: NCC-HN159M [RHH7217]


title: NCC-HN159M [RHH7214]


title: NCC-HN159M [RHH7213]


title: NCC-HN159M [RHH7216]


title: NCC-HN159M [RHH7207]


title: NCC-HN159M [RHH7209]


title: NCC-HN159M [RHH7211]


title: NCC-HN159M [RHH7208]


title: NCC-HN159M [RHH7206]


title: NCC-HN159M [RHH7204]


title: NCC-HN159M [RHH7203]


title: NCC-HN159M [RHH7210]


title: NCC-HN159M [RHH7201]


title: NCC-HN159M [RHH7215]


title: NCC-HN159M [RHH7200]


title: NCC-HN159M [RHH7212]


title: NCC-HN159M [RHH7197]


title: NCC-HN159M [RHH7198]


title: NCC-HN159M [RHH7199]


title: NCC-HN159M [RHH7202]


title: NCC-HN159M [RHH7195]


title: NCC-HN159M [RHH7196]


title: NCC-HN159M [RHH7194]


title: NCC-HN159M [RHH7219]


title: NCC-HN159M [RHH7193]


title: NCC-HN159M [RHH7192]


title: NCC-HN159M [RHH7205]


title: NCC-HN159M [RHH7190]


title: NCC-HN159M [RHH7189]


title: NCC-HN159M [RHH7188]


title: NCC-HN159M [RHH7187]


title: NCC-HN159M [RHH7183]


title: NCC-HN159M [RHH7178]


title: NCC-HN159M [RHH7170]


title: NCC-HN159M [RHH7182]


title: NCC-HN159M [RHH7164]


title: NCC-HN159M [RHH7168]


title: NCC-HN159M [RHH7176]


title: NCC-HN159M [RHH7180]


title: NCC-HN159M [RHH7172]


title: NCC-HN159M [RHH7179]


title: NCC-HN159M [RHH7181]


title: NCC-HN159M [RHH7166]


title: NCC-HN159M [RHH7174]


title: NCC-HN159M [RHH7191]


title: NCC-HN159M [RHH7171]


title: NCC-HN159M [RHH7165]


title: NCC-HN159M [RHH7184]


title: NCC-HN159M [RHH7167]


title: NCC-HN159M [RHH7177]


title: NCC-HN159M [RHH7162]


title: NCC-HN159M [RHH7175]


title: NCC-HN159M [RHH7185]


title: NCC-HN159M [RHH7186]


title: NCC-HN159M [RHH7161]


title: NCC-HN159M [RHH7160]


title: NCC-HN159M [RHH7158]


title: NCC-HN159M [RHH7169]


title: NCC-HN159M [RHH7173]


title: NCC-HN159M [RHH7159]


title: NCC-HN159M [RHH7163]


title: NCC-HN159M [RHH7155]


title: NCC-HN159M [RHH7156]


title: NCC-HN159M [RHH7153]


title: NCC-HN159M [RHH7141]


title: NCC-HN159M [RHH7152]


title: NCC-HN159M [RHH7136]


title: NCC-HN159M [RHH7154]


title: NCC-HN159M [RHH7135]


title: NCC-HN159M [RHH7137]


title: NCC-HN159M [RHH7157]


title: NCC-HN159M [RHH7140]


title: NCC-HN159M [RHH7139]


title: NCC-HN159M [RHH7138]


title: NCC-HN159M [RHH7133]


title: NCC-HN159M [RHH7131]


title: NCC-HN159M [RHH7130]


title: NCC-HN159M [RHH7125]


title: NCC-HN159M [RHH7132]


title: NCC-HN159M [RHH7127]


title: NCC-HN159M [RHH7124]


title: NCC-HN159M [RHH7134]


title: NCC-HN159M [RHH7126]


title: NCC-HN159M [RHH7128]


title: NCC-HN159M [RHH7123]


title: NCC-HN159M [RHH7121]


title: NCC-HN159P [RHT2872]


title: NCC-HN159M [RHH7120]


title: NCC-HN159P [RHT2871]


title: NCC-HN159M [RHH7129]


title: NCC-HN159P [RHT2870]


title: NCC-HN159P [RHT2868]


title: NCC-HN159P [RHT2867]


title: NCC-HN159P [RHT2862]


title: NCC-HN159P [RHT2861]


title: NCC-HN159P [RHT2865]


title: NCC-HN159P [RHT2859]


title: NCC-HN159P [RHT2858]


title: NCC-HN159P [RHT2860]


title: NCC-HN159M [RHH7122]


title: NCC-HN159P [RHT2854]


title: NCC-HN159P [RHT2855]


title: NCC-HN159P [RHT2852]


title: NCC-HN159P [RHT2856]


title: NCC-HN159P [RHT2848]


title: NCC-HN159P [RHT2850]


title: NCC-HN159P [RHT2849]


title: NCC-HN159P [RHT2863]


title: NCC-HN159P [RHT2866]


title: NCC-HN159P [RHT2869]


title: NCC-HN159P [RHT2846]


title: NCC-HN159P [RHT2845]


title: NCC-HN159P [RHT2844]


title: NCC-HN159P [RHT2857]


title: NCC-HN159P [RHT2853]


title: NCC-HN159P [RHT2843]


title: NCC-HN159P [RHT2842]


title: NCC-HN159P [RHT2864]


title: NCC-HN159P [RHT2838]


title: NCC-HN159P [RHT2837]


title: NCC-HN159P [RHT2840]


title: NCC-HN159P [RHT2851]


title: NCC-HN159P [RHT2839]


title: NCC-HN159P [RHT2834]


title: NCC-HN159P [RHT2836]


title: NCC-HN159P [RHT2841]


title: NCC-HN159P [RHT2833]


title: NCC-HN159P [RHT2832]


title: NCC-HN159P [RHT2835]


title: NCC-HN159P [RHT2825]


title: NCC-HN159P [RHT2827]


title: NCC-HN159P [RHT2826]


title: NCC-HN159P [RHT2830]


title: NCC-HN159P [RHT2820]


title: NCC-HN159P [RHT2818]


title: NCC-HN159P [RHT2828]


title: NCC-HN159P [RHT2819]


title: NCC-HN159P [RHT2822]


title: NCC-HN159P [RHT2829]


title: NCC-HN159P [RHT2824]


title: NCC-HN159P [RHT2821]


title: NCC-HN159P [RHT2823]


title: NCC-HN159P [RHT2817]


title: NCC-HN159P [RHT2816]


title: NCC-HN159P [RHT2815]


title: NCC-HN159P [RHT2814]


title: NCC-HN159P [RHT2831]


title: NCC-HN159P [RHT2811]


title: NCC-HN159P [RHT2812]


title: NCC-HN159P [RHT2810]


title: NCC-HN159P [RHT2808]


title: NCC-HN159P [RHT2807]


title: NCC-HN159P [RHT2806]


title: NCC-HN159P [RHT2804]


title: NCC-HN159P [RHT2800]


title: NCC-HN159P [RHT2803]


title: NCC-HN159P [RHT2802]


title: NCC-HN159P [RHT2798]


title: NCC-HN159P [RHT2799]


title: NCC-HN159P [RHT2792]


title: NCC-HN159P [RHT2789]


title: NCC-HN159P [RHT2801]


title: NCC-HN159P [RHT2788]


title: NCC-HN159P [RHT2793]


title: NCC-HN159P [RHT2795]


title: NCC-HN159P [RHT2791]


title: NCC-HN159P [RHT2797]


title: NCC-HN159P [RHT2794]


title: NCC-HN159P [RHT2813]


title: NCC-HN159P [RHT2786]


title: NCC-HN159P [RHT2787]


title: NCC-HN159P [RHT2809]


title: NCC-HN159P [RHT2784]


title: NCC-HN159P [RHT2785]


title: NCC-HN159P [RHT2805]


title: NCC-HN159P [RHT2781]


title: NCC-HN159P [RHT2782]


title: NCC-HN159P [RHT2779]


title: NCC-HN159P [RHT2780]


title: NCC-HN159P [RHT2778]


title: NCC-HN159P [RHT2783]


title: NCC-HN159P [RHT2796]


title: NCC-HN159P [RHT2790]


title: NCC-HN148M [RHC6652]

value: R retromolar trigone

value: Yes Right Level 2


title: NCC-HN148M [RHC6651]


title: NCC-HN159P [RHT2777]


title: NCC-HN148M [RHC6653]


title: NCC-HN148M [RHC6649]


title: NCC-HN148M [RHC6648]


title: NCC-HN148M [RHC6643]


title: NCC-HN148M [RHC6637]


title: NCC-HN148M [RHC6644]


title: NCC-HN148M [RHC6646]


title: NCC-HN148M [RHC6634]


title: NCC-HN148M [RHC6645]


title: NCC-HN148M [RHC6636]


title: NCC-HN148M [RHC6647]


title: NCC-HN148M [RHC6633]


title: NCC-HN148M [RHC6632]


title: NCC-HN148M [RHC6635]


title: NCC-HN148M [RHC6631]


title: NCC-HN148M [RHC6630]


title: NCC-HN148M [RHC6629]


title: NCC-HN148M [RHC6628]


title: NCC-HN148M [RHC6627]


title: NCC-HN148M [RHC6626]


title: NCC-HN148M [RHC6625]


title: NCC-HN148M [RHC6624]


title: NCC-HN148M [RHC6650]


title: NCC-HN148M [RHC6617]


title: NCC-HN148M [RHC6616]


title: NCC-HN148M [RHC6615]


title: NCC-HN148M [RHC6613]


title: NCC-HN148M [RHC6612]


title: NCC-HN148M [RHC6614]


title: NCC-HN148M [RHC6610]


title: NCC-HN148M [RHC6606]


title: NCC-HN148M [RHC6608]


title: NCC-HN148M [RHC6605]


title: NCC-HN148M [RHC6607]


title: NCC-HN148M [RHC6609]


title: NCC-HN148M [RHC6601]


title: NCC-HN148M [RHC6602]


title: NCC-HN148M [RHC6600]


title: NCC-HN148M [RHC6623]


title: NCC-HN148M [RHC6598]


title: NCC-HN148M [RHC6599]


title: NCC-HN148M [RHC6595]


title: NCC-HN148M [RHC6593]


title: NCC-HN148M [RHC6597]


title: NCC-HN148M [RHC6592]


title: NCC-HN148M [RHC6594]


title: NCC-HN148M [RHC6596]


title: NCC-HN148M [RHC6590]


title: NCC-HN148M [RHC6591]


title: NCC-HN148M [RHC6588]


title: NCC-HN148M [RHC6611]


title: NCC-HN148M [RHC6586]


title: NCC-HN148M [RHC6587]


title: NCC-HN148M [RHC6603]


title: NCC-HN148M [RHC6589]


title: NCC-HN148M [RHC6604]


title: NCC-HN148M [RHC6585]


title: NCC-HN148M [RHC6584]


title: NCC-HN148M [RHC6581]


title: NCC-HN148M [RHC6583]


title: NCC-HN148M [RHC6582]


title: NCC-HN148M [RHC6579]


title: NCC-HN148M [RHC6578]


title: NCC-HN148M [RHC6577]


title: NCC-HN148M [RHC6576]


title: NCC-HN148M [RHC6575]


title: NCC-HN148M [RHC6572]


title: NCC-HN148M [RHC6571]


title: NCC-HN148M [RHC6569]


title: NCC-HN148M [RHC6570]


title: NCC-HN148M [RHC6567]


title: NCC-HN148M [RHC6566]


title: NCC-HN148M [RHC6565]


title: NCC-HN148M [RHC6564]


title: NCC-HN148M [RHC6562]


title: NCC-HN148M [RHC6563]


title: NCC-HN148M [RHC6560]


title: NCC-HN148M [RHC6559]


title: NCC-HN148M [RHC6561]


title: NCC-HN148M [RHC6558]


title: NCC-HN148P [RHH2559]


title: NCC-HN148P [RHH2556]


title: NCC-HN148P [RHH2555]


title: NCC-HN148M [RHC6574]


title: NCC-HN148P [RHH2554]


title: NCC-HN148M [RHC6573]


title: NCC-HN148P [RHH2553]


title: NCC-HN148P [RHH2551]


title: NCC-HN148P [RHH2552]


title: NCC-HN148P [RHH2550]


title: NCC-HN148P [RHH2549]


title: NCC-HN148P [RHH2548]


title: NCC-HN148M [RHC6568]


title: NCC-HN148P [RHH2546]


title: NCC-HN148P [RHH2544]


title: NCC-HN159P [RHT2847]


title: NCC-HN148P [RHH2543]


title: NCC-HN148P [RHH2542]


title: NCC-HN148P [RHH2539]


title: NCC-HN148P [RHH2540]


title: NCC-HN148P [RHH2541]


title: NCC-HN148P [RHH2538]


title: NCC-HN148P [RHH2558]


title: NCC-HN148P [RHH2557]


title: NCC-HN148P [RHH2534]


title: NCC-HN148P [RHH2535]


title: NCC-HN148P [RHH2537]


title: NCC-HN148P [RHH2536]


title: NCC-HN148P [RHH2547]


title: NCC-HN148P [RHH2545]


title: NCC-HN148P [RHH2527]


title: NCC-HN148P [RHH2532]


title: NCC-HN148P [RHH2531]


title: NCC-HN148P [RHH2524]


title: NCC-HN148P [RHH2528]


title: NCC-HN148P [RHH2525]


title: NCC-HN148P [RHH2529]


title: NCC-HN148P [RHH2530]


title: NCC-HN148P [RHH2523]


title: NCC-HN148P [RHH2526]


title: NCC-HN148P [RHH2521]


title: NCC-HN148P [RHH2522]


title: NCC-HN148P [RHH2520]


title: NCC-HN148P [RHH2518]


title: NCC-HN148P [RHH2515]


title: NCC-HN148P [RHH2513]


title: NCC-HN148P [RHH2512]


title: NCC-HN148P [RHH2514]


title: NCC-HN148P [RHH2516]


title: NCC-HN148P [RHH2511]


title: NCC-HN148P [RHH2517]


title: NCC-HN148P [RHH2509]


title: NCC-HN148P [RHH2508]


title: NCC-HN148P [RHH2510]


title: NCC-HN148P [RHH2507]


title: NCC-HN148P [RHH2533]


title: NCC-HN148P [RHH2506]


title: NCC-HN148P [RHH2505]


title: NCC-HN148P [RHH2519]


title: NCC-HN148P [RHH2504]


title: NCC-HN148P [RHH2501]


title: NCC-HN148P [RHH2498]


title: NCC-HN148P [RHH2500]


title: NCC-HN148P [RHH2496]


title: NCC-HN148P [RHH2492]


title: NCC-HN148P [RHH2494]


title: NCC-HN148P [RHH2497]


title: NCC-HN148P [RHH2491]


title: NCC-HN148P [RHH2493]


title: NCC-HN148P [RHH2488]


title: NCC-HN148P [RHH2487]


title: NCC-HN148P [RHH2490]


title: NCC-HN148P [RHH2477]


title: NCC-HN148P [RHH2478]


title: NCC-HN148P [RHH2474]


title: NCC-HN148P [RHH2486]


title: NCC-HN148P [RHH2485]


title: NCC-HN148P [RHH2473]


title: NCC-HN148P [RHH2479]


title: NCC-HN148P [RHH2472]


title: NCC-HN148P [RHH2476]


title: NCC-HN148P [RHH2502]


title: NCC-HN148P [RHH2465]


title: NCC-HN148P [RHH2468]


title: NCC-HN148P [RHH2503]


title: NCC-HN148P [RHH2499]


title: NCC-HN148P [RHH2469]


title: NCC-HN148P [RHH2466]


title: NCC-HN148P [RHH2489]


title: NCC-HN148P [RHH2475]


title: NCC-HN148P [RHH2470]


title: NCC-HN137M [RHH1658]

value: L lateral tongue

value: Yes Left Level 2 & Level 3


title: NCC-HN148P [RHH2464]


title: NCC-HN137M [RHH1656]


title: NCC-HN148P [RHH2471]


title: NCC-HN137M [RHH1659]


title: NCC-HN137M [RHH1660]


title: NCC-HN137M [RHH1657]


title: NCC-HN137M [RHH1655]


title: NCC-HN137M [RHH1654]


title: NCC-HN137M [RHH1651]


title: NCC-HN137M [RHH1652]


title: NCC-HN137M [RHH1653]


title: NCC-HN137M [RHH1647]


title: NCC-HN137M [RHH1649]


title: NCC-HN137M [RHH1650]


title: NCC-HN137M [RHH1648]


title: NCC-HN148P [RHH2467]


title: NCC-HN137M [RHH1661]


title: NCC-HN137M [RHH1646]


title: NCC-HN137M [RHH1645]


title: NCC-HN137M [RHH1643]


title: NCC-HN137M [RHH1644]


title: NCC-HN137M [RHH1642]


title: NCC-HN137M [RHH1641]


title: NCC-HN137M [RHH1640]


title: NCC-HN137M [RHH1636]


title: NCC-HN137M [RHH1628]


title: NCC-HN137M [RHH1639]


title: NCC-HN137M [RHH1626]


title: NCC-HN137M [RHH1638]


title: NCC-HN137M [RHH1630]


title: NCC-HN137M [RHH1622]


title: NCC-HN137M [RHH1624]


title: NCC-HN137M [RHH1627]


title: NCC-HN148P [RHH2495]


title: NCC-HN137M [RHH1621]


title: NCC-HN137M [RHH1619]


title: NCC-HN137M [RHH1618]


title: NCC-HN137M [RHH1623]


title: NCC-HN137M [RHH1616]


title: NCC-HN137M [RHH1617]


title: NCC-HN137M [RHH1615]


title: NCC-HN137M [RHH1620]


title: NCC-HN137M [RHH1614]


title: NCC-HN137M [RHH1613]


title: NCC-HN137M [RHH1611]


title: NCC-HN137M [RHH1637]


title: NCC-HN137M [RHH1625]


title: NCC-HN137M [RHH1610]


title: NCC-HN137M [RHH1607]


title: NCC-HN137M [RHH1608]


title: NCC-HN137M [RHH1600]


title: NCC-HN137M [RHH1606]


title: NCC-HN137M [RHH1598]


title: NCC-HN137M [RHH1599]


title: NCC-HN137M [RHH1609]


title: NCC-HN137M [RHH1595]


title: NCC-HN137M [RHH1591]


title: NCC-HN137M [RHH1594]


title: NCC-HN137M [RHH1596]


title: NCC-HN137M [RHH1597]


title: NCC-HN137M [RHH1593]


title: NCC-HN137M [RHH1586]


title: NCC-HN137M [RHH1589]


title: NCC-HN137M [RHH1587]


title: NCC-HN137M [RHH1590]


title: NCC-HN137M [RHH1588]


title: NCC-HN137M [RHH1583]


title: NCC-HN137M [RHH1585]


title: NCC-HN137M [RHH1584]


title: NCC-HN137M [RHH1582]


title: NCC-HN137M [RHH1612]


title: NCC-HN137M [RHH1581]


title: NCC-HN137M [RHH1580]


title: NCC-HN137M [RHH1579]


title: NCC-HN137M [RHH1577]


title: NCC-HN137M [RHH1578]


title: NCC-HN137M [RHH1576]


title: NCC-HN137M [RHH1592]


title: NCC-HN137M [RHH1572]


title: NCC-HN137M [RHH1575]


title: NCC-HN137M [RHH1574]


title: NCC-HN137M [RHH1573]


title: NCC-HN137M [RHH1571]


title: NCC-HN137M [RHH1570]


title: NCC-HN137M [RHH1569]


title: NCC-HN137M [RHH1566]


title: NCC-HN137P [RHH1565]


title: NCC-HN137M [RHH1568]


title: NCC-HN137P [RHH1564]


title: NCC-HN137P [RHH1562]


title: NCC-HN137P [RHH1561]


title: NCC-HN137P [RHH1560]


title: NCC-HN137P [RHH1563]


title: NCC-HN137P [RHH1558]


title: NCC-HN137P [RHH1556]


title: NCC-HN137P [RHH1559]


title: NCC-HN137P [RHH1557]


title: NCC-HN137P [RHH1554]


title: NCC-HN137P [RHH1555]


title: NCC-HN137P [RHH1553]


title: NCC-HN137P [RHH1552]


title: NCC-HN137P [RHH1550]


title: NCC-HN137P [RHH1548]


title: NCC-HN137P [RHH1547]


title: NCC-HN137P [RHH1545]


title: NCC-HN137P [RHH1546]


title: NCC-HN137M [RHH1567]


title: NCC-HN137P [RHH1542]


title: NCC-HN137P [RHH1543]


title: NCC-HN137P [RHH1540]


title: NCC-HN137P [RHH1537]


title: NCC-HN137P [RHH1535]


title: NCC-HN137P [RHH1538]


title: NCC-HN137P [RHH1539]


title: NCC-HN137P [RHH1536]


title: NCC-HN137P [RHH1533]


title: NCC-HN137P [RHH1532]


title: NCC-HN137P [RHH1531]


title: NCC-HN137P [RHH1529]


title: NCC-HN137P [RHH1530]


title: NCC-HN137P [RHH1527]


title: NCC-HN137P [RHH1528]


title: NCC-HN137P [RHH1551]


title: NCC-HN137P [RHH1526]


title: NCC-HN137P [RHH1549]


title: NCC-HN137P [RHH1525]


title: NCC-HN137P [RHH1544]


title: NCC-HN137P [RHH1519]


title: NCC-HN137P [RHH1518]


title: NCC-HN137P [RHH1541]


title: NCC-HN137P [RHH1517]


title: NCC-HN137P [RHH1514]


title: NCC-HN137P [RHH1515]


title: NCC-HN137P [RHH1516]


title: NCC-HN137P [RHH1511]


title: NCC-HN137P [RHH1512]


title: NCC-HN137P [RHH1534]


title: NCC-HN137P [RHH1513]


title: NCC-HN137P [RHH1507]


title: NCC-HN137P [RHH1506]


title: NCC-HN137P [RHH1504]


title: NCC-HN137P [RHH1503]


title: NCC-HN137P [RHH1502]


title: NCC-HN137P [RHH1501]


title: NCC-HN137P [RHH1499]


title: NCC-HN137P [RHH1510]


title: NCC-HN137P [RHH1509]


title: NCC-HN137P [RHH1508]


title: NCC-HN137P [RHH1498]


title: NCC-HN137P [RHH1500]


title: NCC-HN137P [RHH1496]


title: NCC-HN137P [RHH1497]


title: NCC-HN137P [RHH1495]


title: NCC-HN137P [RHH1494]


title: NCC-HN137P [RHH1491]


title: NCC-HN137P [RHH1492]


title: NCC-HN137P [RHH1493]


title: NCC-HN137P [RHH1490]


title: NCC-HN137P [RHH1488]


title: NCC-HN137P [RHH1489]


title: NCC-HN137P [RHH1487]


title: NCC-HN137P [RHH1505]


title: NCC-HN137P [RHH1484]


title: NCC-HN137P [RHH1482]


title: NCC-HN137P [RHH1480]


title: NCC-HN137P [RHH1477]


title: NCC-HN137P [RHH1478]


title: NCC-HN120M [RHH2365]

value: L Tongue

value: Yes Left Level 1 & Level 2


title: NCC-HN137P [RHH1475]


title: NCC-HN120M [RHH2366]


title: NCC-HN137P [RHH1483]


title: NCC-HN137P [RHH1486]


title: NCC-HN137P [RHH1481]


title: NCC-HN137P [RHH1472]


title: NCC-HN137P [RHH1485]


title: NCC-HN137P [RHH1470]


title: NCC-HN137P [RHH1479]


title: NCC-HN137P [RHH1476]


title: NCC-HN120M [RHH2367]


title: NCC-HN120M [RHH2362]


title: NCC-HN137P [RHH1471]


title: NCC-HN120M [RHH2363]


title: NCC-HN120M [RHH2361]


title: NCC-HN120M [RHH2360]


title: NCC-HN120M [RHH2364]


title: NCC-HN120M [RHH2359]


title: NCC-HN120M [RHH2358]


title: NCC-HN137P [RHH1473]


title: NCC-HN137P [RHH1474]


title: NCC-HN120M [RHH2357]


title: NCC-HN120M [RHH2350]


title: NCC-HN120M [RHH2349]


title: NCC-HN120M [RHH2348]


title: NCC-HN120M [RHH2340]


title: NCC-HN120M [RHH2341]


title: NCC-HN120M [RHH2337]


title: NCC-HN120M [RHH2351]


title: NCC-HN120M [RHH2334]


title: NCC-HN120M [RHH2326]


title: NCC-HN120M [RHH2347]


title: NCC-HN120M [RHH2336]


title: NCC-HN120M [RHH2333]


title: NCC-HN120M [RHH2339]


title: NCC-HN120M [RHH2332]


title: NCC-HN120M [RHH2338]


title: NCC-HN120M [RHH2322]


title: NCC-HN120M [RHH2323]


title: NCC-HN120M [RHH2335]


title: NCC-HN120M [RHH2321]


title: NCC-HN120M [RHH2325]


title: NCC-HN120M [RHH2320]


title: NCC-HN120M [RHH2319]


title: NCC-HN137M [RHH1629]


title: NCC-HN120M [RHH2318]


title: NCC-HN120M [RHH2316]


title: NCC-HN120M [RHH2315]


title: NCC-HN120M [RHH2314]


title: NCC-HN120M [RHH2304]


title: NCC-HN120M [RHH2306]


title: NCC-HN120M [RHH2303]


title: NCC-HN120M [RHH2312]


title: NCC-HN120M [RHH2302]


title: NCC-HN120M [RHH2297]


title: NCC-HN120M [RHH2300]


title: NCC-HN120M [RHH2291]


title: NCC-HN120M [RHH2290]


title: NCC-HN120M [RHH2293]


title: NCC-HN120M [RHH2292]


title: NCC-HN120M [RHH2299]


title: NCC-HN120M [RHH2324]


title: NCC-HN120M [RHH2289]


title: NCC-HN120M [RHH2301]


title: NCC-HN120M [RHH2288]


title: NCC-HN120M [RHH2296]


title: NCC-HN120M [RHH2287]


title: NCC-HN120M [RHH2317]


title: NCC-HN120M [RHH2286]


title: NCC-HN120M [RHH2285]


title: NCC-HN120M [RHH2284]


title: NCC-HN120M [RHH2283]


title: NCC-HN120M [RHH2313]


title: NCC-HN120M [RHH2305]


title: NCC-HN120M [RHH2298]


title: NCC-HN120M [RHH2294]


title: NCC-HN120M [RHH2295]


title: NCC-HN120M [RHH2282]


title: NCC-HN120M [RHH2281]


title: NCC-HN120M [RHH2278]


title: NCC-HN120M [RHH2279]


title: NCC-HN120M [RHH2277]


title: NCC-HN120M [RHH2280]


title: NCC-HN120M [RHH2274]


title: NCC-HN120M [RHH2275]


title: NCC-HN120M [RHH2276]


title: NCC-HN120P [RHH2271]


title: NCC-HN120M [RHH2272]


title: NCC-HN120M [RHH2273]


title: NCC-HN120P [RHH2265]


title: NCC-HN120P [RHH2268]


title: NCC-HN120P [RHH2269]


title: NCC-HN120P [RHH2264]


title: NCC-HN120P [RHH2263]


title: NCC-HN120P [RHH2267]


title: NCC-HN120P [RHH2262]


title: NCC-HN120P [RHH2261]


title: NCC-HN120P [RHH2260]


title: NCC-HN120P [RHH2259]


title: NCC-HN120P [RHH2258]


title: NCC-HN120P [RHH2257]


title: NCC-HN120P [RHH2255]


title: NCC-HN120P [RHH2254]


title: NCC-HN120P [RHH2253]


title: NCC-HN120P [RHH2250]


title: NCC-HN120P [RHH2252]


title: NCC-HN120P [RHH2251]


title: NCC-HN120P [RHH2248]


title: NCC-HN120P [RHH2270]


title: NCC-HN120P [RHH2247]


title: NCC-HN120P [RHH2266]


title: NCC-HN120P [RHH2243]


title: NCC-HN120P [RHH2249]


title: NCC-HN120P [RHH2244]


title: NCC-HN120P [RHH2241]


title: NCC-HN120P [RHH2240]


title: NCC-HN120P [RHH2239]


title: NCC-HN120P [RHH2238]


title: NCC-HN120P [RHH2237]


title: NCC-HN120P [RHH2236]


title: NCC-HN120P [RHH2235]


title: NCC-HN120P [RHH2233]


title: NCC-HN120P [RHH2234]