Fig. 7

TNFα-induced Ca²⁺ influx had no effect on mitochondria dependent intrinsic apoptosis. a Typical confocal microscope images (left) of mitochondrial membrane potential using TMRM fluorescent probe in HCC cells treated as indicated. Quantification analysis of mitochondrial membrane potential were shown in the right. Mock: PBS; TNFα: 100 ng/mL. Scale bar: 20 μm. b Cytochrome c release was measured by immunofluorescence in HCC cells with treatment as indicated. Mock: PBS; TNFα: 100 ng/mL. Scale bar: 20 μm. c Western blot analysis for expression of Bcl-2, BAX and BAK in cytoplasm and mitochondria of HCC cells with treatment as indicated. β-actin and COX IV were used as loading controls for cytoplasm and mitochondria, respectively. d Schematic depicting the cytosolic Ca²⁺ signaling on TNFα-mediated cell apoptosis pathway in HCC cells and underlying mechanics. Data were shown as mean ± SD. All experiments were performed at least three times. * P < 0.05; ** P < 0.01