Fig. 4.

GzmB deficiency reduces angiogenic growth factor, VEGF, and macrophages in outer retina. A–C Vascular endothelial growth factor (VEGF) immunofluorescence is robust in outer retina of WT but dramatically reduced in the GzmB−/−. A, B VEGF immunofluorescence (IF) reveals strongly labeled RPE (arrows) and endothelial cells in walls of vessels (arrowheads) in WT compared to GzmB−/−. C Negative control demonstrates the specificity of the IF in which the primary VEGF antibody was omitted and replaced with a non-immune antibody of the same isotype, while keeping all other steps identical during processing. D Average percentage of pixels positive for VEGF immunofluorescence is significantly greater in WT compared to GzmB−/− (n = 4, * p < 0.05, Mann–Whitney U Test). E, F Perivascular macrophage markers F4/80 immuno-fluorescence (green, arrow) is significantly more robust in outer retina of WT vs. GzmB−/−. G Negative control demonstrates the specificity of the IF in which the primary F4/80 antibody was omitted and replaced with a non-immune antibody of the same isotype, while keeping all other steps identical during processing H, I NSE IF (red, arrows), labels macrophages and is significantly more robust in outer retina of WT vs GzmB−/−. J Negative control demonstrates the specificity of the IF in which the NSE reagent was omitted and replaced while keeping all other steps identical during processing. K–L Number of labeled profiles is significantly greater in WT compared to GzmB−/− (n = 4, * = p < 0.05, Mann–Whitney U Test). All tissue samples are from 9-month-old WT or GzmB−/− mice