Increased accumulation of aggregated and truncated α-synuclein in A53T mice. (A) Western blot analysis shows high molecular weight (HMW) α-syn-positive bands in brainstem homogenates of A53T mice as compared to nTg controls. β-actin serves as loading control. (B-C) Western blot analysis shows the presence of HMW α-syn was more abundant in the brainstem than the cerebral cortex of A53T mice at different time points (B). Bar graph shows the ratio of HMW-α-syn/monomeric (Mono)-α-syn in the cortex (Ctx) and brainstem (BS) of A53T mice during the progression of paralysis (C). ***p < 0.0001. (D-E) Western blot analysis shows the level of α-syn in sequentially detergent-extracted cerebral cortex and brainstem homogenates of nTg and A53T mice (D). Bar graph reveals the ratio of total, TX-soluble, and TX-insoluble α-syn in the Ctx and BS of symptomatic A53T mice (E). Signal intensities of α-syn were normalized by β-actin. *p < 0.05. (F-G) Western bolt analysis shows the level of C-terminal truncated α-syn (pointed by arrows) using an antibody that specifically recognizes N-terminal 1-100 amino acids of α-syn (F). Bar graph depicts the ratio of truncated α-syn against total α-syn in the Ctx and BS of symptomatic A53T mice (G). *p < 0.05.