Fig. 2

Subretinal pigment-laden MPs accumulate in CD47^−/−mice with age but not in Thbs1^−/−-mice. Representative micrographs of phalloidin (red fluorescence staining), IBA1 (green fluorescence staining) double-labeled RPE flatmounts of 12-month-old WT (A), Thbs1^−/−- (B) and Cd47^−/−- (C) mice. Asterixis in C represent pigment foci that block the phalloidin fluorescence. Representative bright field- (D) and fluorescence-microscopy (E) views of an anti-IBA1, Hoechst nuclear stain labeled cryo-section of a retinal pigmented focus of a 12-month-old Cd47^−/−-mouse. Quantification of IBA1 stained subretinal MPs of the indicated mouse strains at the indicated ages (F); quantification at 12 months of the percentage of pigment-laden melanophages (that block the Alexa Fluor 594-phalloidin staining of the underlying RPE when viewed in the red channel) of total subretinal MPs (G) and the size of the cell body of the subretinal MP expressed as the surface they cover (H) on flatmounts (n = replicates represent quantifications of eyes from different mice of at least three different experiments and cages; one-way Anova/Kruskal–Wallis test F *p = 0,0028 6 m Cd47^−/−- versus WT-mice; p = 0,0324 6 m Thbs1^−/−- versus WT-mice; *p < 0,0001 12 months Cd47^−/−- versus WT-mice and 12 months Thbs1^−/−- versus WT-mice; *p = 0,0019 18 months Cd47^−/−- versus WT-mice; p = 0,0253 18 months Thbs1^−/−- versus WT-mice; G *p < 0.0001 Cd47^−/−-mice versus WT- and Thbs1^−/−- mice; H *p < 0.0001 Cd47^−/−-mice versus WT- and Thbs1^−/−- mice). Thbs1 Thrombospondin 1 gene, IBA1 ionized calcium-binding adapter molecule 1, ONL outer nuclear layer. Scale bar = 50 µm; All values are reported as mean ± SEM