fig4

Figure 4. IFNγ-sEVs suppress retinal IFN-responsive microglia activation. (A) Expression heatmap of DAM and IRM typical marker genes in all groups; (B) Immunofluorescent staining of IBA1 (red) and LPL (green) in the retinas of ONC mice treated with sEVs at 7 days were shown in representative high-resolution confocal images. Scale bar, 20 μm; (C) The image exhibits the typical expression of LPL⁺ in the IBA1⁺ cells co-located with sEVs in the retinas at a higher magnification. Scale bar, 30 μm; (D and F) Quantification of the number of LPL⁺ in the IBA1⁺ microglia in GCL (D) or IPL (F) (n = 5 eyes per group); (E and G) Quantification of the proportion of LPL⁺ microglia in GCL (E) or IPL (G) (n = 5 eyes per group); (H) Immunofluorescent staining of IBA1 (red) and IFITM3 (green) in the retinas of ONC mice treated with sEVs at 7 days are shown in representative high-resolution confocal images. Scale bar, 20 μm; (I) The image shows the typical expression of LPL⁺ in the IBA1⁺ cells co-located with sEVs in the retinas at a higher magnification. Scale bar, 30 μm; (J and L) Quantification of the number of IFITM3⁺ in the Iba1⁺ microglia in GCL (J) or IPL (L) (n = 5 eyes per group); (K and M) Quantification of the proportion of IFITM3⁺ microglia in GCL (K) or IPL (M) (n = 5 eyes per group). Using one-way ANOVA (D, E, F, G, J, K, L, and M). Data are shown as mean ± SD. ONC: Optic nerve crush; ONL: outer nuclear layer; IPL: inner plexiform layer; LPL: lipoprotein lipase; GCL: ganglion cell layer; IFNγ: interferon-gamma; sEVs: small extracellular vesicles; DAM: disease-associated microglia; IRM: interferon-responsive microglia; IFITM3: interferon-induced transmembrane protein 3; NS: no significant difference. ^*P < 0.05; ^**P < 0.01; ^***P < 0.001; ^****P < 0.0001.