Fig. 4. AMPK participates in the inhibitory effects of EGCG on cardiac hypertrophy in H9C2 cardiomyocytes. H9C2 cardiomyocytes treated with Compound C were pretreated with 25µM EGCG for 1 h followed by stimulation with PE for 24 h. (A) Levels of phospho-/total AMPK were measured by western blotting. (B) Nppa and BNP mRNA levels were measured by quantitative RT-PCR with GAPDH as endogenous control. (C and D) Cardiomyocytes were stained with rhodamine-phalloidin followed by cell surface area quantization. (E) The protein expression of phosphor-/total p70S6K and phosphor-/total eEF2 were detected by western blotting. (F) H9C2 cardiomyocytes were preincubated with a catalase (200 U/ml) or L-NAME (5 mM) for 1 h and subsequently incubated with the indicated concentrations of EGCG for 24 h. Levels of phosphor-/total AMPK were measured by western blotting. (G) Model for EGCG inhibiting cardiac hypertrophy. ^*p<0.05 vs. the group without treatment, ^#p<0.05 vs. the group treated with PE alone, and ^$p<0.05 vs. the group treated with PE plus EGCG, n=3.