Figure 3 | Cell Death & Disease

Figure 3

From: MCL-1 and BCL-xL-dependent resistance to the BCL-2 inhibitor ABT-199 can be overcome by preventing PI3K/AKT/mTOR activation in lymphoid malignancies

Figure 3

BIM associates with increased MCL-1 and BCL-xL protein in ABT199-R cells. MCL-1 and BCL-xL were immunoprecipitated with (a) MCL-1- and (b) BCL-xL-specific antibodies and their association with BIM and NOXA was probed by immunoblotting with specific antibodies. Reciprocal immunoprecipitation-western blotting for BIM was performed in (c) SU-DHL-6 (d) OCL-LY-19 parental and ABT199-R cell lines and its association with MCL-1, BCL-xL, and BCL-2 was analyzed. (e) Parental and resistant cells were treated with ABT-199 at the indicated time and BIM immunoprecipitates were examined by immunoblotting for association with MCL-1, BCL-xL, and BCL-2 in SU-DHL-6 parental and ABT199-R cells. (f) SU-DHL-6 ABT199-R cells were transfected with siAKT, siMcl-1, siBcl-xL or siControl and treated with the indicated concentration of ABT-199 for 24 h. Cell viability was determined by Annexin V-PI staining. Control cells were treated with dimethyl sulfoxide (*P<0.02, **P<0.001) (N=2). β-actin was used as loading control. The experiments from a to e are representative of three independent experiments. L.E and H.E: low and high exposure, respectively. #, non-specific band

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