Journal: PLoS ONE

Article Title: Robo 4 Counteracts Angiogenesis in Herpetic Stromal Keratitis

doi: 10.1371/journal.pone.0141925

Figure Lengend Snippet: WT and Robo4 knockout mice (R4 KO) were infected with HSV-1 RE. (A) R4 KO mice received either sR4 (R4 KO treated w/ sR4) or vehicle (R4 KO Control) from 2 to 14 days pi. WT mice were included and received vehicle (WT Control) under the same regimen previously stated. (B) SK lesions and angiogenesis severity was decreased in R4 KO mice treated with sR4. (C) Representative eye photos show that R4 KO mice treated with sR4 do not develop the severe phenotype that R4 KO control animals do (D) Hematoxylin and eosin staining was carried out on 6-μm sections, and pictures were taken 40 x magnification. Representative eye sections show decreased cellular infiltration in R4 KO treated w/ sR4 and WT control compared to R4 KO control mice. Data are representative of two independent experiments and show mean values ± SEM (n = 12 mice/group). *** p ≤ 0.001,** p ≤ 0.01,* p ≤ 0.05. Statistical levels of significance were analyzed by one-way ANOVA test with Tuckey’s post hoc test settings.

Article Snippet: R4 KO mice were separated into two groups, one of which was treated with sR4 (R4 KO treated) and the other with vehicle (R4 KO control). sR4 was kindly donated by Ryan Watts (Genentech Inc.).

Techniques: Knock-Out, Infection, Control, Staining

Journal: PLoS ONE

Article Title: Robo 4 Counteracts Angiogenesis in Herpetic Stromal Keratitis

doi: 10.1371/journal.pone.0141925

Figure Lengend Snippet: WT mice were infected with HSV-1 RE and treated with sR4 or vehicle (PBS) (A) The sR4 treatment was given to HSV-1 infected mice as shown (B) Dose dependent inhibition of angiogenesis scores after sR4 treatment. Data represents means ± SEM from three different experiments (n = 10) (C) sR4 treatment regimen resulted in SK and angiogenesis scores reduction. Data are representative of three independent experiments and show mean values ± SEM ( n = 15 mice/group). *** p ≤ 0.001, ** p ≤ 0.01, * p ≤ 0.05. Statistical levels of significance were analyzed by t test. (D) Representative eye photos show decreased SK lesion and angiogenesis severity in sR4 treated mice compared to control mice (E) Eyes were processed for cryo-sections on day 15 pi. Hematoxilyn and eosin staining was carried out on 6 μm sections, and pictures were taken at 40x magnification. The sections show decreased cellular infiltration in mice treated with sR4 compared to control mice.

Article Snippet: R4 KO mice were separated into two groups, one of which was treated with sR4 (R4 KO treated) and the other with vehicle (R4 KO control). sR4 was kindly donated by Ryan Watts (Genentech Inc.).

Techniques: Infection, Inhibition, Control, Staining

Journal: PLoS ONE

Article Title: Robo 4 Counteracts Angiogenesis in Herpetic Stromal Keratitis

doi: 10.1371/journal.pone.0141925

Figure Lengend Snippet: WT mice were infected with HSV-1 RE and treated with sR4 of vehicle (PBS) from day 2 pi. to 14 pi. 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 decrease in sR4 treated mice compared to WT 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. (D) Relative fold change in mRNA expression of IL-1β, IL-6 and CXCL-1 was examined and compared between sR4 treated and control mice on day 15 pi. by Q-RT-PCR. Data represent means ± SEM from three different independent experiments (n = 2 and each sample is representative of 6 corneas). *** p ≤ 0.001, ** p ≤ 0.01, * p ≤ 0.05. Statistical levels of significance were analyzed by t test.

Article Snippet: R4 KO mice were separated into two groups, one of which was treated with sR4 (R4 KO treated) and the other with vehicle (R4 KO control). sR4 was kindly donated by Ryan Watts (Genentech Inc.).

Techniques: Infection, Flow Cytometry, Reverse Transcription Polymerase Chain Reaction, Expressing, Control