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Fig. 1 | Journal of Neuroinflammation

Fig. 1

From: Complement activation contributes to subretinal fibrosis through the induction of epithelial-to-mesenchymal transition (EMT) in retinal pigment epithelial cells

Fig. 1

Complement activation in subretinal fibrosis. Subretinal fibrosis was induced in C57BL/6J mice using the two-stage laser-induced protocol (see Methods). A Representative epifluorescence images showing C5b-9 deposition in Col-I+ subretinal lesion in RPE/choroid flatmount counterstained with DAPI. Scale bar = 100 µm. B Schematic diagram showing experimental design of the in vivo C5 blockade study. BB5.1 antibody was administered intraperitoneally on day 0 and 5 after the second laser. Peripheral blood was obtained on days 1, 6 and 10 (end point) for complement activity measurement. Fundus image and fluorescence angiography were carried out immediately before killing the animals on day 10. C Complement activity in the serum of subretinal fibrosis mice treated with control IgG. D Complement activity in the serum of BB5.1 antibody and IgG-treated mice at different times. Data expressed as % of complement activity. Mean ± SEM, n = 3–5 animals. **p < 0.01, one-way ANOVA with Tukey’s multiple comparison tests in C; ****p < 0.001 compared to correspondent vehicle controls by multiple t-test in D. E, F. Representative fundus colour photography (E) and fluorescence angiography (F) in IgG and BB5.1-treated mice at day 10 post second laser. G Quantitative measurement of the area of fluorescein leakage from fibrosis lesion (in pixels). Mean ± SEM, n = 20–21 lesions from 5–6 eyes/group. ***p < 0.005, Student t test. H Representative images of Col-I+ lesions in RPE/choroid flatmount of IgG (left) and BB5.1 (right) treated mice. Scale bar = 100 μm. I Quantitative measurement of Col-I+ lesion area in IgG and BB5.1-treated mice. Mean ± SEM, n = 29–31 lesions from 8 eyes. **p < 0.01; Mann–Whitney test

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