Lentiviral Transduction for Protein Modulation

The pMD-VSV-G and delta 8.2 plasmids were gifts from Dr. Dong at Tulane University. Bax, Bak, and non-target control (NTC) shRNA lentiviral vectors were purchased from Sigma-Aldrich. Precision LentiORF Mcl‐1 and RFP (red fluorescent protein) lentivirus vectors were purchased from Dharmacon (Lafayette, CO, USA). Lentivirus production and transduction were carried out as previously described.^{41 (link)} Briefly, TLA-HEK293T cells were transfected with pMD-VSV-G, delta 8.2, and lentiviral shRNA or LentiORF constructs using Lipofectamine and Plus reagents (Thermo Fisher Scientific) 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 or blasticidin.