(A) Creation of BACHDAlfy mice and the experimental littermates, wild-type (WT), AlfyHet and BACHD.
(B) Representative immunoblot of BACHD (B) and BACHDAlfy (BA) mice probed for Htt (3B5H10) and Alfy. Alfy levels corrected by vinculin (vinc) at right.
(C-F) Depletion of Alfy accelerates accumulation of mHtt in BACHD mice. (C, D) Immunohistochemistry against the EM48 epitope in 12 m/o (C) cortex and (D) striatum. Black arrows indicate cytoplasmic and perinuclear punctate structures; white arrowheads indicate diffuse cytoplasmic or neuritic staining. Scale bar=10 m. (E, F) S830-positive mHtt puncta in the cortex and striatum. (E) S830-positive structures in the cortex of 12 m/o mice. Similar structures were revealed in the striatum (not shown). (F) Stereological quantification of aggregate load. ANOVA revealed a significant increase of aggregates in the BACHDAlfy brain compared to BACHD (Cortex: F(1,11)=15.920, p=0.0021; Striatum: F(1,11)=9.058, p=0.0119), and no significant effect of age (Cortex: F(1,11)=0.242, p=0.6324; Striatum: F(1,11)=0.520, p=0.4857). Data are shown as mean ± SD. Unlike aggregated proteins, detergent soluble proteins are unaffected (See Figure S2).