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. 2016 Jun 16;7(6):e2266. doi: 10.1038/cddis.2016.167

Figure 4.

Figure 4

ABT-737 induces Bax activation in the absence of Bid, Bim, Puma, Bak, and Mcl-1. (a) Diagram for generating HexaKO cells. (b) Western blot of the cell lysates from the wild-type and HexaKO cells. * indicates a non-specific protein. (c) Expression level of GFP-Bax in the indicated cells. Cell lysates were western blotted against an anti-Bax or anti-GFP antibody. (d) HexaKO cells reconstituted with GFP or GFP-Bax were treated with ABT-737 (2.5 μM) for 6 h. The cell lysates were western blotted with anti-PARP antibody. (e) HexaKO cells were treated with ABT-737 (2.5 μM) in the presence of z-VAD (50 μM) for six hours before they were harvested. These cells were lysed in buffer A with 2% CHAPS and loaded onto a Superdex 200 column for gel-filtration analysis. Fractions were western blotted against an anti-GFP antibody. (f) HexaKO cells were treated with ABT-737 in the presence of Z-VAD for 6 h and then stained with MitoTracker and photographed under a fluorescence microscope