Figure 7.

Autophagy protects cells from apoptosis induced by hydrogen peroxide (H₂O₂). (a) The effect of rapamycin, FK506 and FK506 combined with 3-MA on LC3-II and cleaved caspase-3 expression as detected by western blotting. (b) The effect of rapamycin, FK506 and FK506 combined with 3-MA on H2O2-induced apoptosis as determined by TUNEL staining. Cells were incubated with H₂O₂ (150 μM) in the presence of rapamycin (Rapa, 500 nM), FK506 (1 μM) or FK506 (1 μM) combined with 3-MA (5 mM) for 12 h. (c) The effect of ATG7 knockdown on LC3-II and cleaved caspase-3 expression detected by western blotting. (d) The effect of ATG7 knockdown on H₂O₂-induced apoptosis as determined by TUNEL staining. Cells were transfected with negative control or ATG7 siRNA for 48 h and were subsequently subjected to H₂O₂ (150 μM) for 12 h with or without FK506 (1 μM). Scale bar=100 μM, the data for statistical analysis were from three independent experiments and the percentage of TUNEL-positive cells was scored in four non-overlapping microscopic fields in each experiment. Data are presented as the mean±S.E.M. *P<0.05 versus H₂O₂ alone and ^#P<0.05 versus H₂O₂+FK506, ^†P<0.05 versus si-CTRL and ^§P<0.05 versus si-CTRL+FK506