Fig. 4

Proangiogenic phenotype of progranulin deficient macrophages. a Immunofluorescent staining of laser-irradiated RPE-choroid complex from Grn^+/+ and Grn^−/− mice (9–12 weeks) with anti-VEGF-A (white) and anti-CD68 (red) antibodies. Nuclei and CNV are stained with Hoechst 33342 (blue) and FITC-dextran (green). The white dotted line shows the CNV area. Scale bars, 50 μm. b, c Quantification of mean gray value of VEGF-A (b) and CD68 (c) in FITC⁺ and FITC^- area (Grn^+/+, n = 3; and Grn^−/−, n = 4). Data are the means ± SEMs. ^*P < 0.05 vs. Grn^+/+ (Welch’s t-test). d, e PGRN and VEGF-A expression levels in RAW264.7 cells after transfection by siControl or siGrn#1 and exposure to hypoxia. Data are the means ± SEMs (n = 4). ^*P <0.05; ^**P <0.01 vs. siControl; ^#P <0.05; ^##P <0.01 vs. siControl + Hypoxia (one-way ANOVA followed by Tukey’s tests). f, g PGRN and VEGF-A expression levels in Grn^+/+ or Grn^−/− mice-derived peritoneal macrophages after exposure to hypoxia (n = 4). Data are the means ± SEMs. ^*P < 0.05 vs. Grn^+/+ (Welch’s t-test)