Fig. 4. Effect of BHA on H₂O₂-induced apoptosis. (A) Mouse hepatocytes were pretreated with 5 µM BHA or 1,000 µM NAC for 30 min and then incubated with or without 1,000 µM H₂O₂ for 24 h. Cell viability was measured with a CCK-8 assay. Values are expressed as the mean ± SE of three independent experiments with triplicate dishes. ^*p < 0.05 vs. control; ^**p < 0.05 vs. H₂O₂ alone. (B) Cells were pretreated with 5 µM BHA followed by treatment with 1,000 µM H₂O₂ for 24 h. Scale bar = 50 µm. (C) Cells were pretreated with 5 µM BHA and then incubated with 1,000 µM H₂O₂ for 24 h. Cell lysates were subjected to Western blotting to measure the levels of Bax, Bcl-2, and PARP-1 expression. (D) Densities of bands corresponding to Bax and Bcl-2 protein were measured, and the Bax/Bcl-2 ratio was calculated. Values are expressed as the mean ± SE of three independent experiments. ^*p < 0.05 vs. the control , ^**p < 0.05 vs. H₂O₂ alone.