Supplementary Figure 5: Analysis of inclusion pathology and insoluble α-synuclein in A53T mice.

(a) Regardless of lentiviral treatment, α-synuclein inclusion pathology, as detected by Syn505 staining, was abundant in brainstem and absent in SN of A53T mice at 5 mpi. Scale bar, 20 μm. (n = 3 mice per group). (b-d) At 5 mpi, SN from A53T mice was sequentially extracted with buffers containing 1% Triton followed by 2% SDS. Triton-insoluble/SDS-soluble fractions from A53T TH-RREE and A53T CtrlVect contained similar levels of monomeric α-synuclein as assessed by LB509 (b), Syn211 (c), and SNL-4 (d) antibodies. Vimentin (Vim) was used as a loading control, and appears for Syn211 and SNL-4 as the same blot since the same membrane is shown. (LB509, n = 3 mice per group, P = 0.2605, t = 1.310, d.f. = 4; Syn211, n = 6 mice per group except n = 4 mice for CtrlVect, P = 0.7716, t = 0.3003, d.f. = 8; SNL-4, n = 6 mice per group except n = 4 mice for CtrlVect, P = 0.7355, t = 0.3498, d.f. = 8; two-tailed unpaired Student’s t test). Box plots show median, 25th and 75th percentiles, minimum and maximum values.