title: WES of Homo sapiens: T24 GEM resistant
design: T24 UMUC3, and J82 urothelial carcinoma (UC) cells were obtained from the American Type Culture Collection (Rockville, MD, USA). Both cell lines were cultured in RPMI 1640 (containing 11.1 mM glucose and 2 mM L-glutamine) supplemented with 10% fetal bovine serum and penicillin (100 mg/ml) and incubated at 37C in a humidified chamber under 5% CO2 (normoxia). All cells were passaged every 5 days using 0.05% trypsin-EDTA in PBS when they reached a confluency of 75-80%. The cell lines were validated by short tandem repeat (STR) profiling (BEX CO. LTD, Japan). We generated CR cell lines (T24CR and UMUC3CR) by increasing the CDDP concentration to 3 M over 12 months as previously shown6. Likewise, GEM-resistant (GR) UC cell lines (T24GR and UMUC3GR) were generated by increasing the GEM concentration to 3 M over 18 months. These cells were passaged within 1 month of drug exposure to confirm the persistence of CDDP/GEM resistance. Lysates were clarified by using Blood & Cell Culture DNA extraction Kits (QIAGEN, Venlo, Netherlands). The Agilent SureSelect Human All Exon V6 (58 m) Kit (Agilent Technologies, Inc.) was used for the DNA target enrichment.