Fig. 6
Overexpression of endothelial Slc2a1 suppresses aggravated atherosclerosis in Apoe−/−/Prkaa1VEC-KO mice. a Western-blot analysis and quantification data of protein levels of HIF1A and PFKFB3 in HUVECs transfected with siCtrl-Ad-Ctrl, siPRKAA1-Ad-Ctrl, siCtrl-Ad-Slc2a1, and siPRKAA1-Ad-Slc2a1 for 48 h. n = 4. b Simplified schematic of rescue mechanism under overexpressing of Slc2a1. c Representative images and quantification data of flow cytometry analysis of Edu staining in HUVECs transfected with siCtrl-Ad-Ctrl, siPRKAA1-Ad-Ctrl, and siPRKAA1-Ad-Slc2a1 under oscillating flow for 24 h. n = 8. d Quantification data of flow cytometry analysis of Annexin V staining in HUVECs transfected with siCtrl-Ad-Ctrl, siPRKAA1-Ad-Ctrl, and siPRKAA1-Ad-Slc2a1 under oscillating flow for 24 h. n = 8. e Representative images and quantification data of immunofluorescence staining for Slsc2a1 (red) on partially ligated carotid arteries after 48 h transduction of control adenovirus (Ad-Ctrl) and Slc2a1-overexpressing adenovirus (Ad-Slc2a1) in Prkaa1f/f and Prkaa1VEC-KO mice. The endothelium was visualized by CD31 staining (Alexa Fluor-488, green). Nuclei were counterstained with DAPI (blue). Scale bar: 20 µm; n = 6. f Representative images and quantification data of Edu staining on partially ligated carotid arteries 5 days after transduction of control Ad-Ctrl and Ad-Slc2a1 in Apoe−/−/Prkaa1f/f, Apoe−/−/Prkaa1VEC-KO mice. The endothelium was visualized by CD31 staining (Alexa Fluor-488, green). Nuclei were counterstained with DAPI (blue). Scale bar: 20 µm; n = 6. g, i Representative cross-sections and quantification data of partially lighted carotid arteries of transduction of Ad-Ctrl and Ad-Slc2a1 in Apoe−/−/Prkaa1f/f, Apoe−/−/Prkaa1VEC-KO mice stained with Oil Red O. Scale bar: 100 µm; n = 7. h Representative photomicrographs of lesion on whole carotid arteries after transduction of Ad-Ctrl and Ad-Slc2a1 in Apoe−/−/Prkaa1f/f, Apoe−/−/Prkaa1VEC-KO mice. Scale bar: 1 mm; n = 7. All data were expressed as mean ± SEM. Statistical significance was determined by one-way ANOVA followed by Bonferroni test. *p < 0.05 was considered significant, **p < 0.01, ***p < 0.001
