Quantification and statistical analysis

Statistical analyses were performed using GraphPad Prism 9. For quantification of the SC area, 4–8 20X high-power fields were acquired per individual and the CD31⁺ area was measured and averaged. For comparisons of average measurements between two groups, two-tailed, unpaired t test was used. For comparisons of measurements between more than two groups, one-way ANOVA was used with Tukey’s multiple comparisons test. For comparisons of mean values between two groups of mice at multiple IOP levels, two-way ANOVA was used with Šídák’s multiple comparisons test. For quantification of TIE2 expression, 3–8 high-power fields were acquired per individual and mean TIE2 levels were assessed using ImageJ. Statistical analysis was then performed using a nested t test. Statistical analysis for comparison of gene expression between Foxc2^fl/fl and NC-Foxc2^-/- in scRNA-seq violin plot datasets was completed using a Mann–Whitney test. For quantification of sTIE2 concentration, three biological replicates were measured by ELISA. Statistical analysis was then performed using a Kruskal–Wallis test with Dunn’s multiple comparisons. Pathway enrichment analysis on DEGs was performed using Metascape (79) on the KEGG, Canonical Pathways, GO, Reactome, and CORUM databases. P < 0.05 was determined to be statistically significant.