Figure 13. Supplementation of Sema3G ameliorates ischemic retinopathy in the OIR model.

(A and B) Transduction of ECs with AAV in Sema3G^fl/fl and Cdh5-Cre Sema3G^fl/fl OIR mice. Neonatal mice were injected through the retro-orbital sinus with AAV-Control or AAV-Sema3G at P7 and P12. The retinas were analyzed at P19. (C and D) Immunoblots and quantification analysis showed overexpression of Sema3G in the retinas of AAV-Sema3G–injected mice compared with AAV-Control–injected mice (n = 3 independent experiments). (E) IB4 staining of whole-mount retinas from OIR mice infected with AAV-Control or AAV-Sema3G. (F and G) Quantification of the avascular areas and the NVT area at P19 in OIR, related to E (n = 8, 10, 9, and 9 mice for Sema3G^fl/fl + AAV-Control, Sema3G^fl/fl + AAV-Sema3G, Cdh5-Cre Sema3G^fl/fl + AAV-Control and Cdh5-Cre Sema3G^fl/fl + AAV-Sema3G groups, respectively). (H and I) Schematic illustration of the OIR mice treated with intravitreal injections of recombinant Sema3G. At P15, mouse pups were intravitreally injected with 1 μg IgG or recombinant Sema3G. Retinas were analyzed at P17. (J) Entire eye samples of mice were harvested and homogenized, then prepared for immunoblot analysis of total Sema3G protein abundance. (K) IB4 staining of whole-mount retinas from Sema3G^fl/fl and Cdh5-Cre Sema3G^fl/fl OIR mice injected with IgG or recombinant Sema3G. (L and M) Quantification of the avascular area and NVT area at P17 in OIR, related to K (n = 10, 10, 10, and 10 mice for Sema3G^fl/fl + IgG, Sema3G^fl/fl + Sema3G, Cdh5-Cre Sema3G^fl/fl + IgG and Cdh5-Cre Sema3G^fl/fl + Sema3G groups, respectively). Error bars represent mean ± SEM. **P < 0.01; ***P < 0.001; 1-way ANOVA with Tukey’s multiple comparisons test. Scale bars: 1000 μm (E and K).