Fig. 3

Interactions between connexins and eNOS are present at cell-cell interfaces between mouse aortic endothelial cells. Representative images of proximity ligation assays performed with antibodies targeting Cx37 and eNOS or Cx40 and eNOS. a Close proximity of Cx37 and eNOS in WT mouse aortic endothelium. b Cx40 staining to highlight the intercellular gap junctions. c Merge of (a) and (b) highlighting close proximity of eNOS and Cx37 at the gap junctions between endothelial cells. d Close proximity of Cx40 and eNOS in WT endothelium. e Cx37 staining to highlight the intercellular gap junctions. f Merge of (d) and (e) highlighting close proximity of eNOS and Cx40 at the gap junctions between endothelial cells. g-i Control assays revealed that the intense red staining observed in a-c was no longer observed after omitting the eNOS antibody from the proximity ligation assays (g-i) even though cell-cell junctions could clearly be visualized (h-i). j-l: Similar control stainings, which revealed similar results, were performed with the Cx40 antibodies, but not the Cx37 antibodies present during the proximity ligation assays. Scalebar equals 20 μM