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Fig. 1. CaM distribution is heterogeneous and agonist-dependent. (A) A single, live, freshly dissociated smooth-muscle cell loaded with AF-CaM. The arrowhead denotes the position of the nucleus. (B) CaM immunoblot of portal-vein homogenate demonstrating specificity of the antibody used for imaging. (C) Endogenous CaM distribution monitored by indirect immunofluorescence. (top) Control, unstimulated cell. (Inset) Digitally expanded image to illustrate interplaque distance. (middle) 10 minutes, 51 mM KCl PSS. (bottom) 20 minutes, 10 µM DPBA. Arrows point to the location of filamentous structures. Arrowheads denote the position of the nucleus. (D) Quantitation of endogenous CaM distribution. Bars represent the average ratios of surface to core fluorescence ratios (s/c) of fluorescence from 9-21 cells under control conditions or 10 minutes after 10 µM DPBA or 51 mM KCl PSS. (inset) A diagrammatic example of the output from a single line scan in the region of a cell marked with the white line. ***, P<0.001 compared with control; ##, P<0.01 compared with DPBA treatment.

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