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. 2014 Feb;16(2):147–157. doi: 10.1593/neo.131376

Figure 2.

Figure 2

Reduced MCL-1 level is not solely responsible for increased ABT-737 sensitivity. (A) HCT116 and SW620 cells were treated with DMSO equivalent or 2 µM PI-103 for 24 hours, and the level of Bcl-2, BCL-XL, MCL-1, Bax, Bak, Bim, Bid, Puma, Bad, and Noxa was determined by Western blot analysis. (B–D) HCT116 and SW620 cells were transfected with nontargeting siRNA (NT RNAi) or siRNA targeting MCL-1 (MCL-1 RNAi) and plated for experiments 24 hours later. Cells were treated with 2 µM PI-103 for 24 hours, and the effect on levels of MCL-1, pS473 AKT, and GAPDH was determined by Western blot analysis (B). RNAi cells were treated with 2 µM PI-103 or DMSO equivalent and the indicated concentration of ABT-737 for 3 days and processed as in Figure 1A (C). RNAi cells were treated with 2 µMPI-103 and/or 4 µMABT-737 (NT RNAi), 2 µMABT-737 (HCT116 MCL-1 RNAi), or 1 µMABT-737 (SW620 MCL-1 RNAi) for 24 hours. Cells were stained with APC-conjugated annexin V and 7AAD and analyzed by flow cytometry (D). All blots are representative of three independent experiments, and all graphs represent the means of three independent experiments carried out in triplicate (C) or duplicate (D) ± SEM. *P < .05, **P < .01, and ***P < .001 according to two-tailed unpaired t test.