Fig. 3

Treg depletion leads to blood-retinal barrier damage and innate immune cell infiltration in the subretinal space of young and aged mice. A-B Analysis of RPE cell changes. Representative image of phalloidin (A) indicating the geometry of RPE cells. Stars indicate enlarged cells and arrowheads fragmented cytoskeleton (scale bar = 50 µm). Quantification indicating the number of RPE cells with an enlarged morphology per area of RPE/choroid flatmount (B). C-D Determination of the number of CD68⁺ cells in subretinal space. Representative image of CD68 and phalloidin immunostaining in the RPE/choroid flatmount (C) (scale bar = 50 µm) and (D) quantitative analysis of the number of CD68⁺ cells in the subretinal space. E–F Determination of CD68⁺MHCII⁺ cells in the subretinal space. Representative image (E) (scale bar = 50 µm) and quantification (F) of CD68⁺MHCII⁺ innate immune cells in the subretinal space upon Treg depletion and reconstitution in aged mice (scale bar = 50 µm). G Confirmation of the CD68⁺Tmem119⁻ nature of the phagocytes observed in the subretinal space. Arrowheads indicate the scarce presence of CD68⁺Tmem119⁺ cells (scale bar = 100 µm). Data information: A-F, n = 2–5 mice, data presented as mean ± s.e.m. *P < 0.05; ** P < 0.01; ***P < 0.0051; 1-way ANOVA followed by Bonferroni's multiple comparisons test