Fig. 1.
Pyk2 700–841 region recapitulates cytonuclear regulation. a Schematic representation of Pyk2 constructs. b Expression levels of the various GFP-Pyk2 constructs in PC12 cells. Left panel anti-GFP immunoblot of PC12 cells transfected with GFP-Pyk2 (WT), deletion mutants (GFP-Pyk2421–1009, GFP-Pyk21–840, GFP-Pyk21–699, GFP-Pyk21–364, GFP-Pyk2700–841, as indicated) or GFP. Right panel immunoblot with an antibody reacting with the N-terminal region of the protein. GFP-Pyk2 is indicated by an asterisk when visible (i.e. N-terminus present). c Transfected PC12 cells were treated with high K+ (High K+) or control solution for 3 min. GFP fluorescence and nuclei stained with DAPI were analyzed with a fluorescence microscope. d Quantification of the number of cells in c with n ≥ c GFP fluorescence. Values are means + SEM, two-way ANOVA: depolarization effect F (6,42) = 9.93, p < 0.0001, deletion effect F (6.42) = 175.95, p < 0.0001, interaction F (1,42) = 73.71, p < 0.0001. Newman–Keuls test: ***p < 0.001 versus control. °°p < 0.01, °°°p < 0.001 versus WT. Scale bar 5 μm