Fig. 6
Differing intensity of MSA prion infection in astrocytes expressing wt and human mutant α-synuclein (A53T). a Quantification of signal intensity of aggregated α-synuclein inclusions [pSyn (S129)] in Tg(SNCA+/+)Nbm (light blue line), TgM83+/+ (black line), and Tg(SNCA*A53T)Nbm (magenta line) astrocytes exposed to 0.5% MSA35 brain homogenate (pons, dark blue circles). Data are plotted with mean and analyzed by one-way ANOVA followed by Tukey’s multicolumn comparison test: ****, P < 0.0001; **, P = 0.0076. b Quantification of pSyn (S129) inclusions formed in MSA-infected Tg(SNCA+/+)Nbm cultures [size I (< 10 μm), size II (10–50 μm), and size III (> 50 μm)] at 0, 7, 14, and 21 days post-exposure (dpe). Cell count of each group is represented as percentage of total cells, and the data were acquired from 16 randomized fields from two replicate experiments (n = 2). c Graphic representation of same lines of Tg astrocytes as in (b) exposed to 0.5, 0.25, 0.125, and 0.0625% TgM83-passaged MSA and 0.5% TgM83+/+ control littermate brain homogenate. b, c Cultures were immunostained for pSyn (S129) at 0, 7, 14, and 21 dpe, and the signal intensity was normalized by cell count. Data are plotted with mean b (n = 3–6) and c (n = 1). d Representative immunographs of primary astrocytes from (b, c) immunostained for human αSyn (green) and pSyn (S129) (red). Merged channels are shown. Nuclei were stained with DAPI (blue). Scale bars, 20 μm