Fig. 8.

Pfkfb3 overexpression attenuates oxidative stress damage in diabetic mouse retina. A Western blot analysis of PFKFB3 in the whole retina extracts. Nondiabetic C57BL/6 mice (WT) and diabetic mice (DR) received the intravitreal injection of sh-Foxo1, or sh-ctrl as indicated. n = 10. B Human specimens include macular epiretinal membranes from patients without DR (non-DR, n = 6 eyes) and retinal fibrovascular membranes from patients with DR (DR, n = 9 eyes). qPCR was conducted to detect the expression of PFKFB3. C Immunofluorescence staining of PFKFB3 in macular epiretinal membranes from non-DR patients (n = 5 eyes) and retinal fibrovascular membranes (n = 6 eyes) from DR patients. Scale bar, 100 μm. D Western blot analysis of PFKFB3 and γH2AX. Nondiabetic C57BL/6 mice (WT) and diabetic mice (DR) received the intravitreal injection of Ads for Pfkfb3 overexpression (ad-Pfkfb3), or vehicle controls (ad-ctrl) as indicated. n = 10. E Immunofluorescence staining of γH2AX. White arrows indicate γH2AX foci. n = 12. Scale bar, 50 μm. F and G Retinal vascular leakage assessed by EB dye. The confocal analysis of whole-mounted retinas (F, scale bar: 500 μm) and statistical results of spectrophotometrically measured EB extravasation (G) are shown. n = 12. H and I The number of acellular capillaries detected by retinal trypsin digestion. Scale bar, 20 μm. Black arrows indicate acellular capillaries. n = 10. All results are displayed as means ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, for B, unpaired two-tailed Student's t-test, for G and I, one-way ANOVA analysis.