Fig. 7.

CCL2 injections increase inflammatory cells in the retina. The gating strategy for whole retina flow cytometry. All leukocytes are identified from live, single cells as CD45⁺ (A). CD45 + cells are stained with CD11b and Lineage (Lin) gate including CD4 and CD8 (T-cells), B220 (B-cells), NK1.1 (NK cells), SiglecF (eosinophils), Ly6G (neutrophils) to define CD11b⁺Lin⁺ and CD11b⁺Lin^neg cells (B). CD11b⁺Lin^neg cells were gated forward to delineate CD64⁺ and CD64^neg cells (C). From CD64⁺ cells, CD45^dim and CD45^high cells were discriminated (D). Representative plots of CD11b⁺Lin⁺ cells from control and CCL2 groups identified Ly6C⁺Cx3cr1^neg neutrophils (E). Representative plots of CD64^neg cells from control and CCL2 groups defined Ly6C⁺CD45⁺ classical monocytes (F). Representative plots of CD45^dim cells from control and CCL2 groups delineated GFP⁺CD206^neg microglia and GFP^negCD206⁺ resident macrophages (G). Representative plots of CD45^high cells from control and CCL2 groups discriminated Ly6C⁺ macrophages (H). CCL2 treatment increased neutrophil (I), classical monocyte (J), and Ly6C⁺ macrophage infiltration into the retina (M) with no change in microglia or GFP^negCD206⁺ macrophages (K–L). N = 3–4 mice (both eyes) per group from 2 independent experiments that were performed on two separate days from different litters and combined, *p < 0.05, two-tailed unpaired t tests with Welch’s correction