title: RA04_DM_TI.AddRG.Reorder.Sort.bam
organism: Homo sapiens
description: Protocols: Cells were lysed with RLT buffer RASFs were obtained from 5 individual patients and were maintained in the DMEM with 10%FBS and 1% penicillin and streptomycin. Synovial tissues were obtained from patients with RA when they underwent joint replacement surgery or synovectomy. All the patients fulfilled the criteria of the American College of Rheumatology and consent forms were completed by the patients before surgery. RASFs were isolated and cultured as described elsewhere. The experiments used proliferating RASFs (from passage 3 or 4). RASFs were plated to 10cm dish and incubated overnight. Next day, RASFs were treated with 2uM of palbociclib or DMSO. After 24 hours, RASFs were treated with 0.2ng/ml of TNF+IL-1b respectively for 24 hours. RASFs were lysed with RLT buffer RNeasy Plus Mini Kit (QIAGEN) according to the manufacture's protocol 1)mRNA enrichment and purification: Oligo dT Selection to enrich the mRNA (For total RNA extracted from human whole blood, globin mRNA are depleted ); 2)RNA fragmentation and cDNA synthesis (second-strand cDNA synthesis with dUTP instead of dTTP); 3)End repair, add A and adaptor ligation; 4)PCR; 5)Circularization and make DNB; 6) Sequencing on DNBSEQ platform
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2023-01-26
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2023-01-26
attributes.tag: External Id
attributes.value: SAMEA112330120
attributes.tag: INSDC center alias
attributes.value: Department of rheumatology, Tokyo medical and dental university
attributes.tag: INSDC center name
attributes.value: Department of rheumatology, Tokyo medical and dental university
attributes.tag: INSDC first public
attributes.value: 2023-01-26T00:17:06Z
attributes.tag: INSDC last update
attributes.value: 2023-01-26T00:17:06Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-10795:RA04_DM_TI.AddRG.Reorder.Sort.bam
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell_type
attributes.value: fibroblast
attributes.tag: common name
attributes.value: human
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: disease
attributes.value: rheumatoid arthritis
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: individual
attributes.value: RA04
attributes.tag: organism part
attributes.value: synovial joint
attributes.tag: sample name
attributes.value: E-MTAB-10795:RA04_DM_TI.AddRG.Reorder.Sort.bam