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. 2015 Dec 14;291(8):4211–4225. doi: 10.1074/jbc.M115.669978

FIGURE 7.

FIGURE 7.

NA-17-induced mitochondrial accumulation of phospho-p53 conformationally activated Bak by reorganizing the Bak·Bcl-xl complex. A, NA-17 induces conformational Bak activation. Cells were immunostained with antibody for active Bak. Images were acquired using an LSM 710 microscope (magnification, 400×). B, subcellular localization and variation of Bak and Bcl-xl after NA-17 treatment. NCI-H460 cells treated with NA-17 were subjected to subcellular fractionation, and immunoblotting was performed with cytosolic (cytosol) and mitochondrial (mito) fractions. β-Actin and cytochrome c oxidase (cox) IV were used as the cytosolic and mitochondrial marker proteins, respectively. C, phospho-p53 (p-p53) accumulation in mitochondria after NA-17 treatment. NCI-H460 cells treated with NA-17 were subjected to subcellular fractionation, and immunoblotting was performed with cytosolic and mitochondrial fractions. β-Actin and cytochrome c oxidase IV were used as the cytosolic and mitochondrial marker proteins, respectively. D, status of the Bak/Bcl-xl interaction and phospho-p53/Bcl-xl interaction in NCI-H460 cells treated with 5 μm NA-17 for 24 h. **, p < 0.01 versus control (Con). IP, immunoprecipitation.