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Rac1 activity mediated Treg functions to support choroidal neovascularization in a mouse model of age-related macular degeneration (AMD). A mouse model of AMD was established by laser induction, and either a Rac1 inhibitor (1A-116), a neutralizing antibody against IL-10, or a neutralizing antibody against transforming growth factor (TGF)-β1 was administered for intervention. A , Immunofluorescence staining for CD34 in the retina tissues from each experimental group. The microvascular density (MVD) was determined by quantifying the relative CD34+ area in each field (scale bar 200 μm). B , The relative levels of the pro-angiogenic factors (VEGFA and Ang2) in ocular venous blood samples were analyzed by ELISA. C , Flow cytometry analysis of CD25+FoxP3+ Tregs in the CD4+ T cell population from the blood samples. D , ELISA analysis of TGF-β1 and interleukin (IL)-10 levels in the blood samples from each experimental group. Data are reported as means±SD; n=6 animals per group, and 6 independent samples per group were analyzed for each experiment. *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001 (ANOVA).

Journal: Brazilian Journal of Medical and Biological Research

Article Title: Rac1 overexpression promotes Treg-derived cytokines to mediate choroidal neovascularization in wet age-related macular degeneration

doi: 10.1590/1414-431X2024e14187

Figure Lengend Snippet: Rac1 activity mediated Treg functions to support choroidal neovascularization in a mouse model of age-related macular degeneration (AMD). A mouse model of AMD was established by laser induction, and either a Rac1 inhibitor (1A-116), a neutralizing antibody against IL-10, or a neutralizing antibody against transforming growth factor (TGF)-β1 was administered for intervention. A , Immunofluorescence staining for CD34 in the retina tissues from each experimental group. The microvascular density (MVD) was determined by quantifying the relative CD34+ area in each field (scale bar 200 μm). B , The relative levels of the pro-angiogenic factors (VEGFA and Ang2) in ocular venous blood samples were analyzed by ELISA. C , Flow cytometry analysis of CD25+FoxP3+ Tregs in the CD4+ T cell population from the blood samples. D , ELISA analysis of TGF-β1 and interleukin (IL)-10 levels in the blood samples from each experimental group. Data are reported as means±SD; n=6 animals per group, and 6 independent samples per group were analyzed for each experiment. *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001 (ANOVA).

Article Snippet: After the initial de-paraffinization and re-hydration, the antigen retrieval was conducted by heating the sections in the citrate antigen recovery solution (Zeye Biotech, China) at 95°C for 15 min, and peroxidase was inactivated in 3% hydrogen peroxide for 15 min. After rinsing and blocking for 1 h in 5% normal goat serum, primary antibody against CD34 (Cat#: ab81289, 2 μg/mL, Abcam) was applied to label the sections overnight.

Techniques: Activity Assay, Immunofluorescence, Staining, Enzyme-linked Immunosorbent Assay, Flow Cytometry