Lentiviral Transduction of Bak, Bax, CHK1 shRNA

The pMD-VSV-G and delta 8.2 plasmids were gifts from Dr. Dong at Tulane University. Bak, Bax, CHK1, and non-target control (NTC) shRNA lentiviral vectors were purchased from Sigma-Aldrich. Red fluorescent protein (RFP), CHK1, and Mcl-1 cDNA constructs were purchased from Thermo Fisher Scientific Biosciences (Lafayette, CO). Lentivirus production and transduction were carried out as previously described [28 (link)]. Briefly, TLA-HEK293T cells were transfected with pMD-VSV-G, delta 8.2, and lentiviral shRNA constructs using Lipofectamine and Plus reagents (Life Technologies) according to the manufacturer's instructions. Virus containing culture medium was harvested 48 h post-transfection. Cells were transduced overnight using 1 mL of virus supernatant and 4 μg of polybrene and then cultured for an additional 48 h prior to selection with puromycin.