Fig. 4: Regulation of Ang1 and Ang2 expression by Akt1 in VSMCs.

a Aortas were isolated from Akt1^WT and Akt1^∆SMC mice, and the expression of Akt1, Ang1, and Ang2 was verified by western blot, RT–PCR, and qPCR (n = 3) analysis. *P < 0.05, Student’s t test. b Aortas were stained with SM22α (green), Ang1 (red), or Ang2 (red). Images were visualized under a confocal microscope at ×40 magnification. Bar, 50 μm. c The fluorescence intensities of Ang1 (n = 11) and Ang2 (n = 8) were quantified with Student’s t test. *P < 0.05. d MASMCs were isolated from Akt1^WT and Akt1^∆SMC mice, and gene expression was assessed by western blot analysis. Akt1 expression was silenced in VSMCs, and the levels of Ang1 and Ang2 expression were verified by western blotting (e), qPCR (f), and immunocytochemistry g analysis (n = 3). The images were visualized under a confocal microscope at ×40 (Ang2) and ×80 (Ang1) magnification. Bar, 50 μm. *P < 0.05, Student’s t test. h The fluorescence intensities of Ang1 (n = 5–8) and Ang2 (n = 4–6) were quantified with Student’s t test. *P < 0.05. Akt1 was ectopically expressed in VSMCs, and the expression levels of Akt1, Ang1, and Ang2 were assessed by western blotting (i), qPCR (j), and immunocytochemistry k analysis (n = 3). Bar, 50 μm. *P < 0.05, Student’s t test. l The fluorescence intensities of Ang1 (n = 4–6) and Ang2 (n = 5) were quantified with Student’s t test. *P < 0.05. The data are presented as the mean ± SEM.