GSM6329645: OVCAR3, EZH2i; Homo sapiens; RNA-Seq
Cells were maintained at 37°C and 5% CO 2 and cultured in DMEM 1X (Gibco, #11995065) containing 10% FBS (R&amp;D Systems, #S11150) without antibiotics. Cells used in the study were passaged for less than 15 passages. 50 mM stock solution of RAC inhibitor (NSC23766, Sigma #553502) was made in DMSO. 5 mM stock solution of EZH2 inhibitor (GSK126, Biovision #2282) was made in DMSO. For all the experiments using these inhibitors, an equivalent amount of DMSO or RAC1i or EZH2i or combination of both was added to cells and incubated for 48 hrs at 37°C and 5% CO 2. For the collection of cells, trypsin was used to dissociate cells from the culture surface. Cells were resuspended in PBS with a target concentration of 700-1200/ul and with >90% alive. Approximately 10,000 cells per sample were targeted for input to the 10X Genomics  6 Chromium system using the Chromium Next GEM Single Cell 3' Kit v3.1 at the Indiana University School of Medicine (IUSM) Center for Medical Genomics satellite core in Bloomington. The libraries were sequenced at the IUSM Center for Medical Genomics using a NovaSeq 6000 with a NovaSeq S2 reagent kit v1.0 (100 cycles) with approximately 450 million read pairs per sample.