Western blotting

For western blotting, Igrov-1 cells were exposed to pitavastatin or ABT-737 or solvent (DMSO) alone or in combination. After 48 h, cell lysates were prepared as described (18) and total protein was quantified using the Bicinchoninic acid (BCA) protein assay kit (Sigma-Aldrich). Membranes were incubated with rabbit anti-human polyclonal PARP antibody (1:1,000) (#9542), rabbit anti-human polyclonal Bim antibody (1:1,000) (#2819), rabbit anti-human polyclonal Bcl-x_L antibody (1:1,000) (#2762), or rabbit anti-human monoclonal Mcl-1 (D35A5) antibody (1:1,000) (#5453; all from Cell Signaling Technology, Inc., Danvers, MA, USA), and mouse anti-human monoclonal GAPDH antibody (1:5,000) (MAB374; EMD Millipore, Billerica, MA, USA) as a loading control. Bands were quantified using AlphaView SA (ProteinSimple, San Jose, CA, USA) and normalized to GAPDH. The amount of cleaved PARP was expressed as a fraction of uncleaved-PARP, and all other proteins were expressed as a fraction of the solvent control.