2025-08-06
ENA first public
ERC000011
ENA-CHECKLIST
Suzhou Ninth People's Hospital
INSDC center name
public
INSDC status
ERS25302100
SRA accession
E-MTAB-15381:pCREB_GIPRA_1
Submitter Id
ArrayExpress
broker name
primary cell line
cell line
periodontal ligament stem cell
cell type
not collected
collection date
human
common name
Protocols: Human periodontal ligament (PDL) tissues were obtained from five healthy orthodontic donors with informed consent. Immediately after extraction, the teeth were rinsed thoroughly with sterile phosphate-buffered saline (PBS) containing 100 U/mL penicillin and 100 μg/mL streptomycin (Cat. No. 15140122, Thermo Fisher Scientific, USA) to remove blood and debris. PDL tissues were carefully scraped from the middle third of the root surface using a sterile scalpel under aseptic conditions to minimize contamination from gingival and pulp tissues. PDLSCs were primarily cultured within α-MEM supplemented with 15% fetal bovine serum (Thermo Fisher Scientific), and 1 × GlutaMAX (Cat. No. 35050061, Thermo Fisher Scientific).The culture medium for inflammatory PDLSCs were additionally supplemented with 10 ng/mL recombinant human FGF-2 (Cat. No. ab9596, Abcam, USA). 5 × 10^7 PDLSCs were cross-linked with 1% formaldehyde at room temperature for 10 min, and the reaction was quenched with 0.125 M glycine. Chromatin was isolated and sheared to an average size of 200-500 bp under the condition (Power, high intensity; Cycle, 30 s On/30 s Off for 30 cycles; Temperature, 4 °C) using a Bioruptor Pico Sonication Device (Cat. No. B01080010, Hologic Diagenode, Belgium). Immunoprecipitation was carried out overnight at 4 °C using 1 μg IP grade antibodies against p-CREB1 (Cat. No. 81871-1-RR, Proteintech), p-ERK1/2 (Thr202/Tyr204), p-STAT3 (Ser727) (same with Western blot), TCF4 (Cat. No. 22337-1-AP, Proteintech). Next day, the immune complex were added 30 μL Pierce Protein A/G magnetic beads (Cat. No. 88802, Thermo Fisher Scientific) slurry after vortex, and washed, eluted, and reverse cross-linked, followed by DNA purification using a VAHTS Universal Plus DNA Library Prep Kit V4 (Cat. No. ND801-01, Vazyme). ChIP and input DNA were used to prepare sequencing libraries with a NEBNext Ultra II RNA Library Prep Kit for Illumina
description
adult
developmental stage
Malocclusion
disease
wild type genotype
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
periodontal ligament
organism part
Homo sapiens
scientific_name
pCREB_GIPRA_1
title