title: GSM4467063: ST1303_DMSO; Homo sapiens; RNA-Seq
library_construction_protocol: RA-FLS (3 RA and 3 healthy donors) were seeded at 3x10e5/well in a 6-well plate with 2 ml media and incubated overnight for cells to attach. Cells were treated with JQ1+ (1 µM) or DMSO and stimulated with 1 ng/ml IL-1β. After 24 hour incubation, cell media were removed and 600 µl RLT lysis buffer containing β-mercaptoethanol (1%) was added to wells for RNA extraction using RNeasy plus mini kit (Qiagen, Hilden, Germany). RNA was extracted from cell cultures with the Qiagen RNeasy Kit (Qiagen, Hilden, Germany). RNA samples (40 ng) were reverse transcribed and sequencing libraries constructed using NuGen Ovation Universal RNA-Seq System (CAT# 0402-A01) according to the manufacturer's protocol. The resulting sequencing libraries were analyzed using the Caliper LabChip GX and quantified using KAPA qPCR. Libraries were then normalized and pooled in batches of 8. Each pool of 8 samples was clustered and sequenced on an Illumina NextSeq500 instrument using 2x100 bp paired end reads, following the manufacturer's protocols. Each library was loaded to obtain ~80 million reads.
attributes.tag: GEO Accession
attributes.value: GSM4467063