<b>Figure 4</b><b> | </b><b>Inoculation of aSyn PFF-2, but not PFF-1, induce motor deficits within 16 weeks</b>

(A) Schematic of the forelimb grip force test, used to measure muscle strength. Subjects grasped a smooth, triangular pull bar with both forelimbs and the force exerted in Newtons (N) was measured. (B) aSyn PFF-2 but not PFF-1 inoculated mice displayed a significant reduction in forelimb muscle strength at 16 weeks post injection (WPI) relative to 8 (2-way RM ANOVA, Group x Session: main effect of group (F2,72= 4.409, p<0.05), main effect of session (F1,72= 13.51, p<0.001), and significant interaction (F2,72= 6.677, p<0.01), in which Šídák's multiple comparisons revealed to be driven by a significant difference within between 8 and 16 weeks within the PFF-2 group (adj. p<0.0001). (C) Schematic of the accelerating Rota-rod, used to measure motor coordination and motor learning. Subjects were placed on a stationary rod which began to accelerate linearly from 5-36 revolutions per minute over 5 minutes, and latency to fall (s) was calculated automatically. (D) No significant difference between groups was found at 8 WPI (2-way RM ANOVA, Group x Session: main effect of session (F9,675= 15.41, p<0.0001), but no main effect of group (p>0.05). A significant interaction was first observed, (F18, 675= 1.767, p<0.05), but Šídák's multiple comparisons revealed no significant differences between groups (adj. p>0.05)). However, (E) aSyn PFF-2 mice exhibited a shorter latency to fall at 16 WPI relative to aSyn PFF-1 and PBS controls (2-way RM ANOVA, Group x Session: Main effect of group (F2,71= 7.436, p<0.01), main effect of session (F9,639= 3.205, p<0.001), but no interaction (p>0.05). To examine this main effect further, Šídák's multiple comparisons revealed a significant difference between aSyn PFF-2 and PBS controls (adj. p<0.01) and aSyn PFF-2 and PFF-1 (adj. p<0.01)). (F) Schematic of the catwalk XT Gait Analysis, used to measure gait and locomotion. Footprints were captured while subjects voluntarily traversed a glass-plated CatWalk runway, towards a dark goal box at the end. (G) A significant difference between 8 and 16 weeks was revealed in the stride length of aSyn PFF-2 inoculated mice (2-way RM ANOVA, Group x Session: no main effect of group (p>0.05), but a main effect of session (F1,24= 4.493, p<0.05), and significant interaction (F2,24= 3.646, p<0.05), in which Šídák's multiple comparisons revealed a significant difference between timepoints within the aSyn PFF-2 group (adj. p<0.01). (H) A significant difference between 8 and 16 weeks was revealed in the swing speed of aSyn PFF-2 inoculated mice (2-way RM ANOVA, Group x Session: no main effect of group (p>0.05), but a main effect of session (F1,24= 6.537, p<0.05), and significant interaction (F2,24= 4.300, p<0.05) in which Šídák's multiple comparisons revealed a significant difference between timepoints within the aSyn PFF-2 group (adj. p<0.01). (I) A significant difference between 8 and 16 weeks was revealed in the swing speed of aSyn PFF-2 inoculated mice (2-way RM ANOVA, Group x Session: no main effect of group (p>0.05), but a main effect of session (F1,24= 9.232, p<0.01)and significant interaction (F2,24= 4.035, p<0.05) in which Šídák's multiple comparisons revealed a significant difference between timepoints within the aSyn PFF-2 group (adj. p<0.01). Data presented as Mean + SEM, group x session Two-way RM ANOVA, ** p<0.01, **** p<0.0001. Graphics made with Biorender.com