Fig. 5

Dual immunofluorescence detection of phosphorylated alpha-synuclein (p-aSyn, S129) and neuronal nuclei (NeuN) in the GRN of M83^+/- mice. A–C Representative low magnification (10X) images showing p-aSyn (S129) and neuronal nuclei (NeuN) immunofluorescence detection in the GRN/Gi of PBS (in 5A), monomeric aSyn (in 5B) and PFF aSyn (in 5C) injected M83^+/- mice at indicated time-points (DPI, days post-injection). The insets show 63X magnified views of the GRN/Gi, highlighted by the white-bordered square in the 10X views (scale bar = 200 µm). Notice the neuritic and cellular aSyn pathology in monomeric aSyn-injected cohort (DPI-120, in 5B) and the PFF aSyn-injected cohort, which increases over time in the latter (DPI-30 to terminal stage DPI ≥ 108; also see Fig. 6B and S7A). A sparse degree of spontaneous aSyn pathology was also seen in the PBS-injected cohort (in 5A, DPI-120, age 7–8 months). Also see Supplementary Figures S5 (additional brain regions with p-aSyn, S129 positivity), S6-7 (markers of inflammatory gliosis) and S9 (p-aSyn S129 detection in the terminal-stage homozygous M83.^+/+ mice, ≥ DPI-34). Primary antibodies in Fig. 5A–C: p-aSyn (S129)- abcam EP1536Y and NeuN- EMD Millipore A60