Fig. 1.

The CITED2 peptide rescues retinal neovascularization and vaso-obliteration in OIR. (A) Immunofluorescent staining of P17 OIR retinas. OIR mice were intravitreally injected with vehicle (Top), the CITED2 peptide (0.68 ng) (Middle), or the CITED2 APAA peptide (0.68 ng) (Bottom). Retinal whole mounts were stained with GS-lectin. Representative images are shown on the Left; the same images are shown on the Right with NV highlighted in red and VO highlighted in yellow as used for quantification. (Scale bars, 1 mm.) (B) Quantification of the percentage of NV area in whole OIR retinas. (C) Quantification of the percentage of VO area in whole OIR retinas. For B and C, n > 8 per group. P values were calculated using one-way ANOVA with Tukey’s multiple comparisons test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. The data represent at least three independent experiments. (D) Expression levels of proangiogenic genes (Vegfa and Epo) and proinflammatory genes (Tnfa, Ccl2, Ccl3, and Il1b) in P17 retinas. qPCR data are shown for total RNA isolated from the retinas of normoxic mice, vehicle-injected OIR mice, and CITED2 peptide-injected OIR mice (n = 5 per each group). P values were calculated using multiple t tests, *P < 0.05, **P < 0.01, ***P < 0.001. (E) Luciferase activity measured from HEK293 cells transfected with a luciferase reporter gene driven by a promoter containing three copies of the hypoxia response element and treated with the CITED2 peptide or the CITED2 APAA peptide (n = 4 per each group). P values were calculated using a two-tailed unpaired t test. **P < 0.01. For B–E, the mean and SEM are shown in red.