Fig. 4.
Pyk2700–841 cytonuclear localization is regulated by calcineurin. a PC12 cells were transfected with GFP-Pyk2700–841 and treated with high K+ (High K+ 40 mM) or control solution for 3 min in the absence or presence of FK506 (2 μM, 20 min before high K+). GFP fluorescence and nuclei stained with DAPI were analyzed. b Quantification of the number of cells in a with n ≥ c GFP. Values are means + SEM, two-way ANOVA: depolarization effect F (1,14) = 80.92, p < 0.0001, FK506 effect F (1,14) = 5.88, p < 0.05, interaction F (1,14) = 17.49, p < 0.001. Newman–Keuls test: *p < 0.05, ***p < 0.001 versus control. °°°p < 0.001 versus high K+. c Anti-GFP immunoblotting of PC12 cells transfected with GFP-Pyk2700–841, S758A, S762A, T765A, S778A or S788A. d GFP fluorescence and DAPI nuclear staining of PC12 cells transfected as in c and treated with high K+ (High K+ 40 mM) or control solution for 3 min. e Percentage of cells in d with n ≥ c GFP. Values are means + SEM, two-way ANOVA: p < 0.05, mutation effect F (5,12) = 14.33, p < 0.0001, depolarization effect F (1,12) = 72.7, p < 0.0001, interaction F (5,12) = 3.82. Newman–Keuls test: *p < 0.05 versus control. °p < 0.05, °°°p < 0.001 versus 700–841 WT. Scale bar 5 μm