Figure 4.

BAPTA-AM and NAC alleviated gefitinib-induced apoptosis in H4 cells. H4 cells were treated with gefitinib (0 and 40 μM) together, with or without BAPTA-AM (5 μM, A–C), or NAC (2 mM, D–G). Cell viability (24 h) was measured with the alamarBlue assay (A,D). Caspase 3 activity (6 h) was measured with enzymatic assay (B,E). Protein extracts (6 h) were subjected to Western blot with indicated antibodies. Representative blots of three independent experiments and quantitative data are shown (C,F). Intracellular free Ca²⁺ (6 h) was measured with Fura-2 AM fluorescence (G). H4 cells were treated with various concentrations of gefitinib (0–40 μM). Protein extracts (6 h) were subjected to Western blot with indicated antibodies. Representative blots of three independent experiments and quantitative data are shown (H). Protein contents were normalized with corresponding total protein or GAPDH. * p < 0.05 vs. untreated control and # p < 0.05 vs. gefitinib (40 μM), n = 4 (A,B,D,E,G).