Figure 3. Treadmill reduces α-synucleinopathy in aged A53T mice brain.

(A–H) Effect of regular treadmill on α-syn aggregates in aged (10 months old) A53T NR and R mice brain was demonstrated by assessing total Triton X-100 soluble and insoluble α-syn in midbrain tissues (F_{2, 11} = 29.44, p < 0.05 for insoluble α-syn, A–D, n = 5) and by double immunostaining of total α-syn in TH-positive neurons of SN coupled with quantification of size, area, and integrated density of α-syn intraneuronal puncta (**p < 0.01, E–H, n = 6). Two sections from each brain per group were used for staining purposes, where α-syn puncta was quantified in at least 10 TH-positive cells from each section.

(I and J) Level of pathological pSyn129 in nigral sections were shown by immunostaining followed by optical density measurement (***p < 0.001, n = 4).

(K and L) Effect of treadmill exercise on motor behavior was monitored by rotarod test and by counting the number of rearing in the open-field test (F_{2, 15} = 6.983, p < 0.05 for rotarod, F_{2, 15} = 24.58, p < 0.01 for rearing, n = 6).

(M–O) Spatial cognitive function of experimental animals was done by Barnes maze, where the latency time to reach goal box and number of errors made by each animal before reaching goal box were counted (F_{2, 12} = 12.35, p < 0.05, n = 5).

Statistical analyses were conducted by unpaired two-tailed t test for comparison between two samples and by one-way ANOVA followed by Tukey’s multiple comparison tests. *p < 0.05, **p < 0.01, ***p < 0.001. Data are represented as mean ± SEM.