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. 2016 Mar 4;7:10830. doi: 10.1038/ncomms10830

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Double immunofluorescent staining for PTEN (green) and αSMA (red) was conducted on arterial tissues from human patients. (a) Confocal microscopic images of human coronary arterial media from two patients (#1 and 2); bottom panels are higher magnification images of boxed areas. Stains are shown separately rather than in overlay to demonstrate both cytoplasmic and nuclear PTEN staining, but exclusive cytoplasmic αSMA expression in normal medial SMCs. (b) Confocal microscopic images of right coronary artery with large atherosclerotic plaque (lower magnification H&E image is shown in Supplementary Fig. 7c; ‘*' and ‘**' represent the region of vessel shown in the H&E image). Top panels demonstrate cytoplasmic and nuclear PTEN staining in medial SMCs. Middle and bottom panels demonstrate loss of nuclear PTEN associated with loss of αSMA expression in plaque intimal SMCs (arrows). Right panels show merged images plus DAPI staining for cell nuclei. Scale bars for all images, 50 μm. H&E, haematoxylin and eosin.