GSM4101949: I-BET151 IFNg Rep2; Homo sapiens; RNA-Seq
All SLAM-seq assays were performed at 60% of the maximum cell density counted on a hemocytometer for suspension cells. 5-7h prior to each assay, growth medium was aspirated and replaced. Newly synthesized RNA was labeled for the last 60 minutes at the end of the treatment with the inhibitors at a final concentration of 100μM 4-Thiouridine. Suspension cells were spun down and immediately snap-frozen, before RNA extraction using the RNeasy Plus Mini Kit (Qiagen). Total RNA was subjected to alkylation by iodoacetamide (Sigma, 10mM) for 15 minutes and RNA was re-purified by ethanol precipitation. Libraries were prepared using QuantSeq 3' mRNA-seq Library Prep kit (Lexigen)