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. 2005 Mar 14;168(6):887–897. doi: 10.1083/jcb.200408128

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Copyright © 2005, The Rockefeller University Press

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Figure 3. — Effect of stimulation on CaMKII phosphorylation and MEF2 reporter activity. (A) Mean nuclear pixel fluorescence (normalized to control) in resting fibers and fibers repeatedly stimulated for 1 h using 10-Hz trains in the presence or absence of KN-62 and labeled with antibody for either autophosphorylated (open bars) or total (closed bars) CaMKII. Only nuclei stimulated in the absence of KN-62 and stained for autophosphorylated CaMKII exhibited a significant increase in nuclear staining after stimulation. From left to right, the data were averages of mean nuclear fluorescence from 26, 17, 23, 21, or 13 nuclei from 22, 12, 18, 12, or 10 fibers, respectively. (B) Luciferase activity was increased in fibers infected with adenovirus-encoding MEF2-luciferase reporter and stimulated with 10-Hz trains for 5 h. 5 μM KN-62 blocked the increase in luciferase activity stimulated with 10-Hz trains. Results represent triplicate measurements from each of three independent experiments. Error bars represent ± 1 SEM. *, P < 0.05 versus control without stimulation.