Fig. 2.

H₂O₂-promoted eNOS phosphorylation and translocation in cardiac myocytes. (A) Results of immunohistochemical analyses of cardiac myocytes that were treated with H₂O₂ (10 μM) for the indicated times, and then fixed, permeabilized, probed with antibodies against total eNOS, peNOS Ser¹¹⁷⁷, or peNOS Ser⁶³³, and imaged using confocal microscopy. (B) Images of cardiac myocytes treated with H₂O₂ (10 μM) for the indicated times and then fixed, permeabilized, and probed with antibodies against total eNOS (Alexa Fluor red 568) or Cav-3 (Alexa Fluor green 488); overlapping signals are shown in yellow. The images shown on the left are representative of three independent experiments that yielded similar results; the bar graph on the right shows pooled data from three experiments, quantitating the percent overlap between eNOS and Cav-3 at different times after adding H₂O₂; * indicates p < 0.05; ** indicates p < 0.01 compared to t = 0.