Kv7.4 channels colocalize with caveolin-1. (A) Representative midcell z section of an isolated smooth muscle cell from a rat small mesenteric artery stained with antibodies against caveolin-1 (Cav-1; magenta) and Kv7.4 (green). Scale bar, 10 µm. Insets show the brightfield images of the cells. (B i) Representative images of PLAs in smooth muscle cells from rat mesenteric arteries with Kv7.4 and caveolin-1 antibodies in control (left) and M-βCD–treated (right) cells. Red puncta indicate target proteins are in close proximity (<40 nM). (B ii) Quantification of the number of PLA puncta in mesenteric artery myocytes (control: 27 cells, n = 3 rats; M-βCD: 29 cells, n = 3 rats) showing significant decrease of Kv7.4 and caveolin-1 colocalization in M-βCD–treated cells (**, P = 0.008 according to an unpaired t test). (C i) Representative images of PLAs in smooth muscle cells from rat mesenteric arteries with NCX and caveolin-1 antibodies in control (left) and M-βCD–treated (right) cells. Red puncta indicate target proteins are in close proximity (<40 nM). (C ii) Quantification of the number of PLA puncta in mesenteric artery myocytes (control, 10 cells; M-βCD, 10 cells) showing equal colocalization of NCX and caveolin-1 in M-βCD–treated cells compared with nontreated cells. Error bars represent SEM.