Fig. 3: Defects in EC proliferation and mural cell coverage in Akt1∆SMC mice.

a Retinas were isolated from Akt1^WT, Akt1^∆SMC, Akt2^WT, and Akt2^∆SMC mice at P6 and stained with IB4 (green) and pH3 (red). White arrowheads indicate IB4- and pH3-positive ECs, and purple arrowheads indicate pH3-positive mural cells. Bar, 100 μm. b Quantification of the number of pH3-positive ECs (n = 5–8). *P < 0.05, one-way ANOVA followed by Tukey’s multiple comparison test. c Aortas were embedded in growth factor-reduced Matrigel-coated plates in the presence of EGM-2. After 7 days, bright field images were captured under an optical microscope at ×5 magnification. d Quantification of the sprouting area (n = 3–4). *P < 0.05, one-way ANOVA followed by Tukey’s multiple comparison test. e, g Retinas were isolated from Akt1^WT and Akt1^∆SMC mice at P4 and stained with IB4 (blue), NG2, or SM22α (red). The images were visualized under a confocal microscope at ×40 (top) and ×80 (bottom) magnification. Bar, 50 μm. f, h The fluorescence intensity of NG2/IB4 (n = 7–9) and SM22α/IB4 (n = 6) was quantified with Student’s t test. *P < 0.05. The data are presented as the mean ± SEM.