Veliparib

Human liver cancer cell lines including PLC/PRF/5, QGY7703, Huh7, etc., normal liver cell line Chang and mouse liver cancer cell line Hepa1-6 were all purchased from Cell Bank of the Typical Culture Preservation Committee, Chinese Academy of Sciences (Shanghai, China). PLC/PRF/5, Huh7,Hepa1-6, HepG2, SK-Hep1 and HCCLM3 cells were cultured in the DMEM medium (Invitrogen, Shanghai, China).And Chang, QGY7703, BEL7402 were cultured in RPMI1640 medium (Invitrogen, Shanghai, China). All mediums were supplemented with 10% FBS and 100U/mL penicillin–streptomycin. The following antibodies were used for Western blotting: WIP1 (sc-376257) to detect human samples from Santa Cruz (Shanghai, China); WIP1 (A6204) to detect mouse samples from ABcolonal (Wuhan, China); cleaved PARP1(#9541), cleaved-caspase3 (#9661) and beta-Actin (#4970) from Cell Signaling Technology (Shanghai, China); γH2AX (phospho-Histone H2AX (s139)) (ab81299) from Abcam (Shanghai, China); ki67 (ER1802-31) from Huabio (Hangzhou, China). WIP1 plasmid was kindly provided by Prof. Zhenyu Ju at Hangzhou Normal University. GSK2830371, Diethylnitrosamine (DEN) and TCPOBOP were purchased from Sigma-Aldrich (Shanghai, China). Olaparib (HY-10162) and Veliparib (HY-10129) were purchased from MedChemExpress (Shanghai, China). Other reagents and chemicals don’t list here are commercially available.

UM-UC-3, T-24 (human bladder cancer cell lines), and SV-HUC-1 (normal human bladder epithelial cell line) were obtained from the American Type Culture Collection (ATCC, Manassas, VA) and were cultured in EMEM, McCoy’s 5a, or F12K media respectively, supplemented with 10% FBS and penicillin/streptomycin. Cells were used within half a year after being received from ATCC or after thawing from cryopreservation. Short Tandem Repeat (STR) profiling is used by the ATCC for cell line authentication. The cell lines in culture were routinely tested for mycoplasma contamination every two months using the MycoFluor^TM Mycoplasma detection kit (Thermo Fisher Scientific, Waltham, MA). Tubulin antibodies were obtained from Thermo Fisher Scientific, Waltham, MA. Cleaved and whole caspases 3, 7, and 9 antibodies were obtained from Cell Signaling Technology, Danvers, MA. Ki-67 antibodies were obtained from Neomarker, Fremont, CA. The PARP inhibitors, olaparib, niraparib, rucaparib, veliparib, and talazoparib, were obtained from MedChem Express, Monmouth Junction, NJ. Cisplatin was from Sigma Aldrich, St. Louis, MO. Other reagents were supplied by local suppliers such as Fisher Scientific and VWR International.