Fig. 7

Anti-Sema3A treatment in primary microglia increased M2-like microglia and decreased RGCs apoptosis in primary microglia and RGC co-culture. a Schematic diagram of primary microglia and RGC co-culture system. b Representative confocal images showing immunofluorescent staining of CD206 (green) and CD16/32 (red) with Hoechst (blue) nuclear staining in Sema3A treated primary microglia with or without anti-Sema3A treatment. Scale bar = 100 μm. Quantitative analysis indicates a significant increase of CD206 with anti-Sema3A treatment. (Mean ± SEM, n = 6). c TUNEL staining of RGCs in control, Sema3A + anti-Sema3A, Sema3A, and LPS groups. Scale bar = 100 μm. d Quantification of TUNEL-positive cells vs. Hoechst staining (n = 4). e–h Western Blot and quantitative analysis show expression of SMI32, MAP2, pro-caspase3, and cleaved-caspase3 in RGCs co-cultured with microglia. β-actin was used as a loading control. (Mean ± SEM, n = 5)