AngII effects on TRPV4 sparklet activity in smooth muscle depend on sex and vascular bed. (A–C) TIRF images of representative male (left) and female (right) pial (A), parenchymal (B), and mesenteric (C) myocytes in the absence (control) and presence of 100 nM AngII. Traces to the right of each image show the time course of [Ca²⁺]_i in regions highlighted by green circles. (D) Amplitude histograms of amalgamated TRPV4 sparklets in male (left) and female (right) myocytes (i.e., pial, parenchymal, and mesenteric myocytes) before (white bars) and after 100 nM AngII (gray bars). Black (control) and red (AngII) lines show the best fits of the male and female data with a multi-Gaussian function with a quantal unit of Ca²⁺ influx of 50 nM. (E and F) Bar plot of mean TRPV4 sparklet activity ± SEM (E) and density ± SEM (F) in male (n = 6–7 cells per condition, 11 animals) and female (n = 8–9 cells per condition, 15 animals) pial, parenchymal, and mesenteric myocytes. *, P < 0.05; **, P < 0.01; ***, P < 0.001.