Figure 2. GLUT1 deficiency leads to early BBB breakdown in Slc2a1^+/− and Slc2a1^+/−APP^Sw/0 mice.

(a) Representative confocal microscopy analysis of fibrin (red) and lectin-positive capillaries (white) in 2-week-old Slc2a1^+/+,Slc2a1^+/−, Slc2a1^+/+APP^Sw/0 and Slc2a1^+/−APP^Sw/0 mice. Scale bar, 25 μm. (b-c) Quantification of extravascular fibrin (b) and immunoglobulin G (IgG) (c) deposits in 2-week old Slc2a1^+/+, Slc2a1^+/−, Slc2a1^+/+APP^Sw/0 and Slc2a1^+/−APP^Sw/0 mice. Mean ± SEM, n=3-5 mice per group; *p<0.05 or **p<0.01. (d-f) Representative immunoblotting of fibrin and IgG levels in capillary-depleted brain tissue (d) and quantification of IgG (e) and fibrin (f) relative abundance in capillary-depleted brains in 2-week-old Slc2a1^+/+, Slc2a1^+/−, Slc2a1^+/+APP^Sw/0 and Slc2a1^+/−APP^Sw/0 mice. β-actin was used as a loading control. Mean ± SEM, n=3-5 mice per group; **p<0.01. (g-i) The tight junction proteins occludin or zonula occludens-1 (ZO-1) (green) and lectin-positive capillary profiles (red) (g) and quantification of endothelial occludin (h) and ZO-1 (i) length in the microvessels in 2-week-old Slc2a1^+/+, Slc2a1^+/−, Slc2a1^+/+APP^Sw/0 and Slc2a1^+/−APP^Sw/0 mouse hippocampus. Scale bar, 25 μm. Mean ± SEM, n=3-5 mice per group; *p<0.05 or **p<0.01. (j-k) Negative correlation between the extracellular fibrin (j) or IgG deposits (k) and decreased occludin length in the microvessels in 2-week-old Slc2a1^+/+, Slc2a1^+/−, Slc2a1^+/+APP^Sw/0 and Slc2a1^+/−APP^Sw/0 mouse hippocampus. (l) Representative immunoblotting analysis of microvascular ZO-1, occludin and claudin-5 (l) and their respective protein abundance in microvessels (m) in a 2-week-old Slc2a1^+/+, Slc2a1^+/−, Slc2a1^+/+APP^Sw/0 and Slc2a1^+/−APP^Sw/0 mouse. β-actin was used as loading control. Mean ± SEM, n=3-5 mice per group; *p<0.05 or **p<0.01. Full length blots are presented in Supplementary Figure 11.