GSM8745660: iPSC_DMSO_2; Homo sapiens; RNA-Seq
Four hours after treatment, cells were washed twice with PBS then treated with 0.25% trypsin-EDTA solution (Sigma-Aldrich, T4049) and transcription/translation inhibitors: 5 μg/mL actinomycin D (Sigma-Aldrich, A1410), 10 μM triptolide (Sigma-Aldrich, T3652), and 27.1 μg/mL anisomycin (Sigma-Aldrich, A9789) for 6 minutes at 37 °C in a 5% CO2 incubator. Then, 1 volume of PBS with 2% FBS (Gibco, A38400-01), the transcription/translation inhibitors, and 1:100 DNAse (Worthington, LS002007) was added to quench. Cells from two wells were combined for each technical replicate and put through 40 μm cell strainers (BD Falcon, 352235). Cells were counted and spun at 4 °C for 5 minutes at 1500 rpm. Supernatant was removed and cells were resuspended at 1,000 cells/μL in ice-cold PBS. Cells were loaded onto the 10x Genomics Next GEM Single Cell 3′ v3.1 sample chip and run following the manufacturerʼs protocols. 10x Genomics Next GEM Single Cell 3′ v3.1