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- _2 value "Activation of nuclear CaMKII and MEF2 To test for changes in CaMKII phosphorylation, fibers were stimulated with a 5-s duration 10-Hz train every 50 s for 1 h with or without 5 microM KN-62 in the Ringer's solution and fixed and stained with primary antibody to activated (autophosphorylated; Fig. 3 A, open bars) or total (Fig. 3 A, filled bars) CaMKII. 1 h of 10-Hz train stimulation significantly increased (P less than 0.01) the mean nuclear stain of activated CaMKII compared with unstimulated fibers, and KN-62 (5 microM) blocked the increase in activated CaMKII staining after stimulation (Fig. 3 A). In parallel experiments, in which the fibers were stained with antibody directed against total CaMKII, there was no difference in mean nuclear antibody stain between stimulated and unstimulated fibers (P greater than 0.05; Fig. 3 A). These results are consistent with activation of nuclear CaMKII by fiber electrical stimulation without any appreciable translocation of CaMKII from the cytoplasm to the nucleus." provenance.