Chronic infusion of AngII increases AKAP150–TRPV4 distance in pial and parenchymal myocytes. (A) Two-color super-resolution localization maps of AKAP150 (red) and TRPV4 (green) in male (left) and female (right) pial and parenchymal myocytes from mice infused with AngII for 7 d. To the right, a zoomed-out image of the region of interest (top) and the binary image (mask, bottom) used for nearest neighbor analysis. The bar plot shows the mean ± SEM distance between AKAP150 and TRPV4 clusters in male (n = 1,000 ROIs from five cells per condition) and female (n = 1,000 ROIs from five cells per condition) pial and parenchymal myocytes from control mice (i.e., saline infusion) and mice chronically infused with AngII. (B) Bar plot of the mean ± SEM cluster density for AKAP150 and TRPV4 in pial myocytes from male and female mice infused with saline solution (control) or with AngII. (C) Real-time PCR analysis of the mean ± SEM fold change of mRNA for AKAP150, TRPV4, and PKCα in control and AngII-infused arteries (n = 3). (D and E) TIRF images of male parenchymal myocytes in control conditions and during acute application of 100 nM AngII. The cell in D was isolated from a mouse infused with saline, whereas the cell in E was isolated from a mouse chronically infused with AngII. Traces at right show the time course of [Ca²⁺]_i in regions highlighted by green circles. (F) The bar plot shows mean ± SEM TRPV4 nP_s values in male parenchymal myocytes (n = 4 cells per treatment, eight animals) from control and AngII-infused mice before and after acute AngII treatment. *, P < 0.05; **, P < 0.01; ***, P < 0.001.