Figure 5.

PKA increases AMPK activity in VSMCs. Cells were treated with FSK (10 uM), PKI (10 uM), or a combination of FSK and PKI or IBMX (10 uM) or AIC (1 mM) for 60 min. Phosphorylation of AMPK Thr172 and ACC Ser80 were measured by In-Cell Western (A) and (B) and AMPK activity was measured by pAMPK-specific activity assay (C). FSK alone failed to significantly increase AMPK Thr172 or ACC Ser80 phosphorylation or AMPK activity; however, in the presence of IBMX or AICAR all were significantly elevated. PKI alone significantly reduced both pAMPK Thr172 and activity. Data are presented as phosphorylated AMPK/total AMPK and normalized to DNA content (Draq 5/Sapphire 700) or absorbance at 540 nm for activity. P-values less than 0.05 were considered statistically significant for n = 5–7 per group. ^*p < 0.05 compared to control; ^***p < 0.001 compared to control.