Fig. 2

Artesunate increases PICALM levels in brain capillaries, slows development of Aβ pathology, increases brain-to-blood clearance of Aβ, and improves functional outcome in Picalm^+/−; 5XFAD mice. A Experimental treatment paradigm for Picalm^+/−; 5XFAD mice with artesunate (Art) or vehicle. Mice were injected i.p. with 32 mg/kg/day Art or vehicle for 2 months, starting at 3 months of age, followed by behavior tests, evaluation of cerebral blood flow (CBF) responses, and tissue collection and assays at 5 months of age. B PICALM protein relative expression levels in brain capillaries isolated from Picalm^+/−; 5XFAD mice treated with vehicle or Art. n = 4 mice per condition. C, D Amyloid-β 42 (Aβ42) (C) and Aβ40 (D) levels in the hippocampus (Hpp) and cortex (Ctx) in Picalm^+/−; 5XFAD mice treated with vehicle or Art. n = 8–10 mice per condition (C: Hpp and Ctx Vehicle: 5 male, 5 female; Hpp and Ctx Art: 5 male, 4 female. D: Hpp Vehicle: 4 male, 5 male; Hpp Art: 4 male, 4 female; Ctx Vehicle: 4 male, 4 female; Ctx Art: 5 male, 4 female). E, F Representative images of Aβ immunostaining (E), and quantification of Aβ load in the Hpp and Ctx (F) in Picalm^+/−; 5XFAD mice treated with vehicle or Art. Scale bar in (E) is 500 μm. n = 10 mice per condition (Hpp and Ctx Vehicle: 5 male, 5 female; Hpp and Ctx Art: 6 male, 4 female). G, H Representative images of Thioflavin S-positive amyloid deposits (G) and quantification of Thioflavin S amyloid plaque load in the Hpp and Ctx (H) in Picalm^+/−; 5XFAD mice treated with vehicle or Art. Scale bar in (G) is 500 μm. n = 9–10 mice per condition (Hpp and Ctx Vehicle: 5 male, 5 female; Hpp Art: 6 male, 4 female; Ctx Art: 6 male, 3 female). I, J Representative images (I) and quantification (J) of Aβ vascular load in small pial arteries and penetrating cortical arterioles in Picalm^+/−; 5XFAD mice treated with vehicle or Art. Aβ vascular load is expressed as a fraction of Aβ-positive area relative to α-smooth muscle actin (αSMA)-positive area of the vessel wall, normalized to the vehicle-treated group. Scale bar in (I) is 25 μm. n = 7 mice per condition. K, L Serum Aβ42 (K) and Aβ40 (L) levels in Picalm^+/−; 5XFAD mice treated with vehicle or Art. n = 8 mice per condition (4 male, 4 female). M Cerebral blood flow (CBF) response to whisker stimulation in in Picalm^+/−; 5XFAD mice treated with vehicle or Art. n = 8–9 mice per condition (Vehicle: 4 male, 4 female; Art: 6 male, 3 female). N, O Representative images (N) and quantification (O) of blood-derived fibrin/fibrinogen extravascular leakage in the brain parenchyma in Picalm^+/−; 5XFAD mice treated with vehicle or Art. Scale bar in (N) is 25 μm. n = 7 mice per condition. P-S Novel object location (P) and recognition (Q), burrowing (R), and nesting score (S) in Picalm^+/−; 5XFAD mice treated with vehicle or Art. n = 9–10 mice per condition (P, R, and S: Vehicle: 5 male, 5 female and Art: 6 male, 4 female; Q: Vehicle: 4 male, 5 female and Art: 6 male, 3 female). Single points per mouse indicated by circles in B-D, F, H, J-M, O, P-S, with boxes representing mean ± SD. Significance determined by Student’s two-tail t-test in panels B, F, H, K-M, R, and S, or Mann-Whitney U test in panels C, D, K, O-Q. Full blots for B shown in supp. Fig. 5