Intersections of kinase pathways and [Ca²⁺]_i in control of H₂O₂-promoted eNOS responses.

<p>In panel A, adult mouse cardiac myocytes were loaded with the NO dye Cu₂(FL2E), and then treated with the AMPK inhibitor Compound C (1 µM) or vehicle followed by hydrogen peroxide (H₂O₂, 10 µM) treatment. Upper panel shows representative fluorescence images at 0, 2, and 5 minutes followed treatments as indicated. Middle panel shows representative fluorescence tracings of a cell treated with H₂O₂ or H₂O₂ in the presence of compound C. Lower panel shows the results of pooled data analyzed from at least three independent repetitions with a minimum of 4 cells analyzed per experiment that yielded equivalent results; *indicates p<0.05. Panels B, C, and D show representative experiments analyzing the effects of nifedipine (100 µM), BAPTA AM, (60 µM), or calphostin C (1 µM), on H₂O₂-promoted AMPK or ACC phosphorylation. Cardiac myocytes were pre-incubated with these compounds for 30 min, then treated with H₂O₂ (25 µM, 30 min) and analyzed in immunoblots probed with phospho-AMPK Thr172, phospho-ACC Ser79, AMPK, or ACC antibodies, as shown. Each data point represents the mean ± S.E. derived from at least three independent experiments; *indicates p<0.05 (<i>ANOVA</i>).