Fig. 1: Medin co-localizes with amyloid-β deposits and promotes vascular β-amyloidosis in mouse models.
From: Medin co-aggregates with vascular amyloid-β in Alzheimer’s disease
a, Schematic of MFG-E8 protein domains in Mfge8 wild-type (WT) and C2 domain knockout (C2 KO) mice and the binding sites of the anti-mouse MFG-E8 antibody (green; dotted and solid lines indicate weak and strong antibody affinity, respectively). TM, transmembrane domain; β-gal, β-galactosidase reporter gene . b, Schematic of pathology and timing of analyses (arrows) in APP transgenic lines. c,d, Immunostaining of cortical brain sections of 4-month-old APPPS1 × Mfge8 wild-type or APPPS1 × Mfge8 C2 KO mice (c) and 27-month-old APP Dutch mice (n = 2 females analysed) and 24-month-old APP23 mice (d, left), with quantification of co-localization (3 female and 3 male 12- to 13-month-old mice, and 2 female and 5 male 21- to 24-month-old mice; total of n = 323 vessels and n = 386 plaques). c, Bottom, reconstructed confocal z-stack. Aβ, amyloid-β. e, Plaque load in Mfge8 wild-type and C2 KO mice in APPPS1 (2-month-old: 7 female and 8 male wild-type mice, 6 female and 9 male C2 KO mice; 4-month-old: 8 female and 8 male wild-type mice, 12 female and 8 male C2 KO mice) and APP23 lines (6-month-old: 8 female wild-type and 8 female C2 KO mice, males have no plaques yet; 9-month-old: 7 female and 7 male wild-type mice, 7 female and 6 male C2 KO mice; 12-month-old: 4 female and 9 male wild-type mice, 6 female and 7 male C2 KO mice; 24-month-old: 4 male wild-type mice, 4 male C2 KO mice). f,g, CAA-laden vessels in 12- and 24-month-old APP23 animals (f) and microhaemorrhages in 24-month-old APP23 animals (g) (numbers of mice as in e). h, Confocal z-stack of an isolated cerebral blood vessel and western blotting for vascular markers (α-smooth muscle actin, α-SMA; platelet-derived growth factor receptor-β, PDGFR-β) and MFG-E8. i, Quantification of MFG-E8 by ELISA (4-month-old APPPS1: 3 female and 3 male mice; 6-month-old APP23: 3 female and 3 male mice; 12- to 13-month-old APP23: 7 female and 7 male mice; 21- to 24-month-old APP23: 2 female and 10 male mice). j, Quantification of total vascular amyloid-β by ELISA (Mfge8 wild-type as in i; APPPS1 × Mfge8 C2 KO: 2 female and 3 male mice; 6-month-old APP23 × Mfge8 C2 KO: 3 female and 3 male mice; 12- to 13-month-old APP23 × Mfge8 C2 KO: 4 female and 4 male mice; 21- to 24-month-old APP23 × Mfge8 C2 KO: 3 male mice). Data are mean ± s.e.m. e,f, right, i,j, Two-way-ANOVA with Tukey’s post hoc comparison. d,f, left, g, Two-tailed Mann–Whitney U-test. Scale bars: 100 µm (c, overview), 20 µm (c, magnified, h), 5 µm (c, z-stack) and 50 µm (d). ND, not detectable; NS, not significant. Uncropped western blots are shown in Supplementary Fig. 1.
