Figure 1.

The effect of hydrogen peroxide (H₂O₂) on cardiomyocyte apoptosis, calcineurin activity, AMPK phosphorylation and LC3 expression. Primary cardiomyocytes were treated with H₂O₂ at a final concentration of 150 μM for 1, 3, 6 or 12 h. (a) The effect of H₂O₂ on cell viability at various time points with or without zVAD-fmk. (b) H₂O₂ induces time-dependent activation of caspase-3 as shown by western blotting. The statistical bar graph (right panel) shows the comparison of cleaved caspase-3 levels that have been normalised to β-actin. (c) H₂O₂ time dependently enhances calcineurin (Cn) activity. Cells were treated as described. Calcineurin activity in each group was normalised to the control group. (d) The effect of H₂O₂ on LC3 and AMPK protein expression at each time point as detected by western blotting. The levels of LC3-II and p-AMPK were normalised to β-actin and are shown in the bar graph (right panel). Data are presented as the mean±S.E.M., n≥3. *P<0.05 versus the control group, ^#P<0.05 versus 3 h and ^†P<0.05 versus H₂O₂ alone at 6 and 12 h