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Fig. 4. UNC-43 calcium-dependent kinase activity. (A) S2 cells were transfected with the indicated cDNAs, then lysed and immunoblotted using anti-V5 antibodies to detect the recombinant proteins. (B) S2 cell lysates made from the indicated transfected cDNAs (squares for rat CaMKII and filled triangles for UNC-43) were tested for their ability to phosphorylate AC-2 in the presence of varying concentrations of calcium. The data were normalized to the maximal activity. (C) AC-2 kinase activities for S2 cell lysates from cells expressing the indicated cDNAs. White bars indicate the activity in the presence of calcium, whereas gray bars indicate the calcium-independent activity in the presence of EGTA. Error bars indicate the s.e.m. Significant differences in kinase activity were found *P<0.05 and ***P<0.001 by one-way ANOVA followed by Bonferoni comparisons between the same transgenes (calcium versus EGTA). n=3 or more trials. (D) The calcium-independent activity (AC-2 kinase activity in the presence of EGTA, plotted as a percentage of the activity in the presence of calcium) of S2 lysates expressing the indicated form of UNC-43. Error bars indicate s.e.m. Significant differences in activity were observed *P<0.05 and ***P<0.001, compared to levels in the WT group by one-way ANOVA followed by Bonferoni comparisons. n=3 or more trials.
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