Fig. 2 |. Treatment of mice with αCXCR2 antisera, starting on the day of i.o. zymosan injection, skews eye-infiltrating neutrophils towards an immature phenotype.

a-c Mice were injected i.o. with zymosan on day 0 of ONC injury, and i.p. with either αCXCR2 (red) or NRS (blue) on days 0, 2 and 4. Inflammatory cells were isolated from the vitreous on day 5. a, b Surface expression of myeloid cell markers. Upper panels, representative histograms. Lower panels, geometic Mean Fluorescence Intensity of Ly6G (n=5 mice/group), CD14 (n=5 mice/group), Ly6B (n=5 mice/group), MPO and elastase on Ly6G⁺ gated cells ( (n=3 mice/group), and % of Ly6G⁺ gated cells that are CD101⁺(n=8 mice/group). Each symbol represents data obtained from a single mouse. One experiment representative of 3 is shown. Statistical significance determined by two tailed unpaired Student’s t-test. b, left panels, Cytospins of purified vitreal Ly6G⁺ cells, Wright Giemsa stained (top panels scale bar 10 μm, bottom panels scale bar 6 μm). Arrows point to granules. c, Mean length of the longest neurite grown by primary RGC, following 24 h co-culture with intraocular Ly6G⁺ neutrophils (N_ϕ) that were purified from the NRS or αCXCR2 treatment groups. RGC were cultured with recombinant ciliary neurotrophic factor (CNTF) as a positive control, or with particulate zymosan (Zym) or media alone (No Tx) as negative controls (n= 200 RGCs per condition). Statistical significance was determined by one-way ANOVA followed by Tukey’s post hoc test. Right panels, representative images, scale bar 20 μm. a, b, c, Data are shown as mean +/− sem.