Robo4 deficient mice present more inflammation and vascularization.

Robo4 knockout (R4 KO) and WT mice were infected with HSV-1 RE and at day 15 pi. corneas were collected and pooled for analysis by flow cytometry or Q-RT-PCR. The frequency and total cell number per cornea for (A) endothelial cells (CD31+) gated on total CD45- cells infiltrate, (B) CD4+ T cells (CD4+) (gated on total CD45+ cells infiltrate) and (C) neutrophils (Ly6G+ CD11b+ gated on total CD45+ cells infiltrate) show significant increase in R4 KO mice. Data are a combination of 3 independent experiments and show mean values ± SEM (n = 7 and each sample is representative of 2 corneas). ***p ≤ 0.001, **p ≤ 0.01, *p ≤ 0.05. Statistical levels of significance were analyzed by t test. (E) Relative fold change in mRNA expression of IL-1β, IL-6 and CXCL-1 was examined and compared between WT and Robo4 KO mice on day 15 pi. by Q-RT-PCR. Data represent means ± SEM from two different independent experiments (n = 3 and each sample is representative of 5 corneas). ***p ≤ 0.001, **p ≤ 0.01, *p ≤ 0.05. Statistical levels of significance were analyzed by t test.