Dynein colocalizes with Kv7.4 proteins in vascular smooth muscle cells. (A) Representative Western blot showing dynein (∼74 kD) expression in protein lysates from three different samples of rat small mesenteric artery (rMA) and in one protein lysate from untransfected HEK293B cells. (B) Representative structured illumination microscopy images of an isolated smooth muscle cell from a rat small mesenteric artery, stained with antibodies against β-tubulin (green) and dynein (magenta). Scale bar, 10 µm. (C) Representative Kv7.4 bands from a Western blot following coimmunoprecipitation of dynein in HEK293B cells overexpressing Kv7.4. No bands were detected for the nonspecific binding sample (NS) or for the negative control sample (Neg ctr), where normal mouse control IgG was used instead of the specific pulldown antibody. (D) Representative Kv7.4 bands from a Western blot with samples of rat mesenteric artery protein lysate (rMA) that were immunoprecipitated (IP) with a dynein antibody. Each rMA sample contains n = 3 rats’ worth of mesenteric arteries. (E) Representative structured illumination microscopy image of an isolated smooth muscle cell from a rat small mesenteric artery stained with antibodies against Kv7.4 (magenta) and dynein (green). Scale bar, 10 µm. (F i) Representative image of PLAs with Kv7.4 and dynein antibodies in mesenteric artery myocytes. Each red punctum is representative of the Kv7 channel and dynein protein localizing within 40 nm of one another. (F ii) Quantification of puncta in 34 cells (n = 4 rats) with Kv7.4 and dynein and the respective antibodies alone as control. Mean values are shown with error bars depicting the SEM.