Fig. 1

The number of mitovesicles in the brain of Ts2 mice is higher than controls. (A) Western blot analyses of EVs isolated from brains of 2N and Ts2 mice. BH: brain homogenate. KDa: kilodaltons. (B-C) Cryo-EM photomicrographs (B) and diameter (C) of Fr8 EVs (mitovesicle-enriched; ‘mtV’) from 2N (n = 148 EVs, 3 mice) and Ts2 (n = 214 EVs, 3 mice) brains. Fr1 EVs (microvesicle-enriched; n = 184 EVs, 3 mice) are the controls. Scale bar: 200 nm. Data are shown as a violin plot. Fr1 EVs vs. mitovesicles P < 0.0001; 2N vs. Ts2 mitovesicles P = 0.1713. Kruskal-Wallis H test by ranks with Dunn’s multiple comparisons test. (D) NTA diameter of mitovesicles from the brain of 2N (n = 14) and Ts2 (n = 19) mice. Distributions normalized to the mode. Trendline: four-point moving average. Genotype P = 0.6271. In (D-G), bars are mean ± SEM and the statistical test used was ordinary two-way ANOVA with Bonferroni’s multiple comparisons test. (E, F) Hydrodynamic size analyses of mitovesicles isolated from 2N (n = 8 males, 9 females) and Ts2 (n = 13 males, 8 females) brains. In (E), for the mean, genotype P = 0.8430, sex P = 0.6148, genotype X sex P = 0.1957, 2N vs. Ts2 within males P = 0.5421, 2N vs. Ts2 within females P = 0.9016; for the median, genotype P = 0.5832, sex P = 0.5298, genotype X sex P = 0.2900, 2N vs. Ts2 within males P = 0.4773, 2N vs. Ts2 within females P > 0.9999. In (F), genotype P = 0.9970, 2N vs. Ts2 for each bin: P > 0.9999. (G) Mitovesicle number isolated from 2N (n = 8 males, 9 females) and Ts2 (n = 12 males, 10 females) brains, quantified by NTA. Genotype P < 0.0001, sex P = 0.6170, genotype X sex P = 0.7450, 2N vs. Ts2 within males P = 0.0027, 2N vs. Ts2 within females P = 0.0098. ** P < 0.01, **** P < 0.0001