FIGURE 3.

17β-Estradiol promotes recruitment of PKGIα but not PKGIβ to the plasma membrane. Isolated hepatocytes seeded onto coverslips were incubated for 5 min at 37 °C in the presence of either no treatment (Ai and Bi), 10 nm 17β-estradiol (Aii and Bii), 10 nm 17β-estradiol plus 25 μm Rp-8-pCPT-cGMPS (Rp) (Aiii and Biii) 10 nm 17β-estradiol plus 4 μm DT-3 (Aiv and Biv), 10 nm 17β-estradiol-BSA (Av), or 10 nm 17α-estradiol (Avi). Cells were then immunostained with either anti-PKGIα (cGKIα (N-16)) (Ai–Avi) or anti-PKGIβ (cGKIβ (L-16)) (Bi–Biv). Alexa-594 anti-goat (red) was used to visualize immunoreactivity. The images and their corresponding intensity scans (Ai–vi and Bi–iv) are representative of several experiments. The scale bar represents 10 μm. Mean percentage staining intensities at the cell periphery (white bars) and the cell interior (gray bars) (as a percentage of average staining intensity across the entire cell transect) were calculated from fluorescence intensity scans of several cells from at least three independent hepatocyte preparations immunostained with either anti-PKGIα (C) or anti-PKGIβ (D).