Fig 2: Influence of sDLL4 on macrophages in CNV in vivo. Representative image of an IBA1-stained choroidal flatmount 4 days after laser-injury of PBS treated mice (a) or sDLL4 treated mice (b). Quantification of IBA1 positive cells per impact in PBS (white, n = 25 impacts) and sDLL4 (black, n = 25 impacts) treated mice (c). Il-1ß (d) and VEGF (e) mRNA levels and VEGF protein concentrations (f) in choroid 4 days following CNV and 24 h after PBS or sDLL4 injections (PBS, white, n = 6 choroid, sDLL4 black, n = 6 choroid). * p < 0.001. Fluorescent activated cell sorting (FACS) for CD11b and F4/80 (g) and LY6B.2 (h) on choroidal cell suspensions 4 days after laser-injury and 24 h after PBS (white columns i–l) or sDLL4 (black columns i–l) treatment. VEGF (j) and Il-1ß (i) RT-PCR on sorted CD11b+F4/80+LY6B.2neg resident macrophages and CD11b+F4/80+ LY6B.2pos inflammatory macrophages. VEGF (l) and IL-1ß (k) RT-PCR on whole choroidal cell suspension and CD11b+F4/80+depleted cell suspension. Choroidal cell suspensions were prepared from 8 choroids per group. The results are representative of two independent experiments. Double labeling of VEGF (green) and F4/80 (red) on choroidal flatmounts of PBS (m) and sDLL4 (n) treated CNV 4 days after laser-impact. Double labeling was performed on n = 3 eyes/group. Arrows indicate F4/80 positive cells. Scale bar 50 µm. (Color figure online)