CD11b⁺ mononuclear phagocytes, CD11c⁺ dendritic cells, endothelial cells and retinal vimentin⁺ cells contain VEGF.

Abstract

<p>(A) VEGF⁺ cells were analyzed for CD11b and CD11c expression. CD11b⁺ CD11c^– phagocytes and CD11b^– CD11c⁺ dendritic cells (DCs) are shown in the choroid and the retina of untreated eyes (left) and 3 days after laser injury (right). (B) The VEGF⁺CD11b⁺ phagocytes were subclassified by analyzing their CD45 and CX₃CR1 expression at day 3. (C) VEGF⁺CD11b⁺ cells in the lasered choroid were further characterized by analyzing F4/80 expression (bottom), which is present on MP but absent on neutrophils. (D) VEGF⁺ endothelial cells (ECs) were identified by CD31 staining in eyes from healthy mice (left) and after laser at day 3 (right) in the choroid (top) and the retina (bottom). (E) Retinal VEGF⁺ cells were identified as mentioned in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0094313#pone-0094313-g001" target="_blank">Figure 1</a> and displayed (top). VEGF⁺ cells were stained positive for vimentin (black line in Histogram) versus the control (grey tinted). (F) Gating strategy to exclude CD31⁺ ECs (1) and CD11b⁺ and CD11c⁺ phagocytes (2) from the vim⁺ cell subset of astrocytes and Müller cells (3). This subset was analyzed for VEGF expression compared to the isotype control. (G) RPE cells were stained for the marker RPE65 (right) versus control (left) in the choroid (top) and the retina (bottom). Percentages (A–D) are means of the respective group. Results are representative for two independent experiments. A total of 3–5 mice per group were analyzed in each experiment.</p