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. 2013 Aug 1;4(8):e744. doi: 10.1038/cddis.2013.252

Figure 4.

Figure 4

Wip1 inhibits radiation-activated BAX mitochondrial translocation. (a) HeLa and LNCaP cells were irradiated (IR) with 10 Gy. After 48 h, the cells were fixed and double-fluorescence stained with MitoTracker (red) for the mitochondria and anti-BAX (green). 4′,6-Diamidino-2-phenylindole (DAPI) counter-stain (blue) was used to label the nuclei. (b) Mitochondrial fractions were extracted from HeLa or LNCaP cells in the presence or absence of CT007093 after 48 h of 10 Gy IR. Mitochondrial proteins or whole-cell lysates (WCL) were subjected to immunoblotting (IB) using anti-BAX, prohibitin (a mitochondrial marker), and aldolase (a cytoplasmic marker). (c) LNCaP cells were irradiated with 10 Gy in the presence or absence of CT007093, followed by the preparation of total cell extracts at the indicated times. BAX proteins were immunoprecipitated (IP) using anti-BAX (6A7) antibody, which recognizes only the active form of BAX, and then analyzed by BAX immunoblotting. Anti-IgG antibody was utilized as a negative control