Additional file 3: of TNFα induces Ca2+ influx to accelerate extrinsic apoptosis in hepatocellular carcinoma cells

Figure S2. Decreased cytosolic Ca2+ level attenuates TNFα-induced apoptosis of HCC cells. (a) Confocal microscope analysis of [Ca2+]c level using fluorescent probe Fura-2/AM in SNU739 and HLF cells with treatment as indicated. BAPTA-AM: 10 μM. (b) Cell apoptosis analysis by flow cytometry 24 h after treatment as indicated before TNFα (100 ng/mL) stimulation. BAPTA-AM: 10 μM. (c) Confocal microscope analysis of [Ca2+]c level using fluorescent probe Fura-2/AM in HCC cells with treatment as indicated. EV: cells transfected with the empty vector; PV-OE: cells stably forced expressing Parvalbumin protein. (d) Apoptosis analysis by flow cytometry 24 h after treatment as indicated. (e) Western Blot analysis for Parvalbumin expression in SNU739 and HLF cells with treatment as indicated. (f) Cell apoptosis analysis by flow cytometry 24 h after treatment as indicated. CAI: 10 μM; SKF96365: 100 μM. (g) Cell apoptosis analysis by flow cytometry 24 h after treatment as indicated. siTRPM7: siRNA against TRPM7; si Ctrl: negative control siRNA. Data were shown as mean ± SD. All experiments were performed at least three times. * P < 0.05; ** P < 0.01. (ZIP 1986 kb)