Fig. 7

Biochemical effect of VPS35 overexpression on human WT-α-Syn levels and phosphorylation in rat brain. A) Western blot analysis of rat ventral midbrain and striatal extracts at 14 weeks post-injection of AAV vectors expressing WT-α-Syn and VPS35 (WT or D620N) or empty control (MCS). 1% Triton-X100 or 2% SDS fractions were probed with antibodies to human VPS35 (V5), human α-Syn (Syn211 antibody), pS129-α-Syn, TH, or β-tubulin as a control. Note, only human WT-α-Syn is detected in the 2% SDS fraction from ventral midbrain or striatum, but pS129-α-Syn or human VPS35 are not detectable. B-C) Densitometric analysis of pS129-α-Syn levels normalized to human α-Syn, or human α-Syn or VPS35 (V5) levels normalized to β-tubulin in 1% Triton-soluble extracts from (B) ventral midbrain or (C) striatum. Bars represent mean ± SEM (n = 4 animals/group). *P < 0.05 by one-way ANOVA with Dunnett’s post-hoc analysis, as indicated. n.s., non-significant