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Fig. 6 | Journal of Neuroinflammation

Fig. 6

From: Progranulin deficiency in Iba-1+ myeloid cells exacerbates choroidal neovascularization by perturbation of lysosomal function and abnormal inflammation

Fig. 6

Lysosomal abnormality in PGRN-deficient macrophages. a Representative images of LysoTracker Red DND-99 staining (red) and phase contrast images of RAW264.7 cells after transfection by siControl or siGrn. Scale bar: 10 μm. b Quantitative analysis of the fluorescence intensity of LysoTracker+ cells. Data are the means ± SEM (n = 6). *P <0.05 vs. siControl (Welch’s t-test). c Representative images of western blots showing immunoreactivity against LAMP1, cathepsin D, and β-actin. df Quantitative analysis of expression level of LAMP1 (d), pre-cathepsin D (e), and mature-cathepsin D (f) in RAW264.7 cells after transfection of siControl or siGrn and exposure to hypoxia (1% O2, 12 h). 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). g, h immunohistochemistry on the laser-injured eye of Grn+/+ and Grn−/− mice 7 days after the photocoagulation with anti-LAMP1 (white) and anti-cathepsin D (magenta) antibodies. Nuclei were stained with Hoechst 33342 (blue) and the CNV regions were visualized by FITC-dextran (green). Yellow arrows show LAMP1+ or cathepsin D+ cells infiltrated into the CNV area. Scale bar: 100 μm. i, j Level of expression of sortilin in RAW264.7 cells after transfection by siControl or siGrn and exposure to hypoxia. Data are the means ± SEMs (n = 4). *P <0.05; **P <0.01 vs. siControl (one-way ANOVA followed by Tukey’s tests)

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