Fig. 5: Role of EV-loaded miRNAs in age-related dysfunction.
a, Bar plots show the different miRNA expression in YBMM-EVs and ABMM-EVs. The yellow and green shading represents the candidate miRNAs, namely miRNA-378a-3p (yellow shading) and miRNA-191 (green shading); the green and red horizontal dashed lines represent the expression levels of miRNA-378a-3p in ABMM-EVs (red dashed line) and miRNA-191 in YBMM-EVs (green dashed line). b, Outline of the studies (2 months old, male mice). c–e, Fasting blood glucose (n = 6 mice) and fasting serum insulin level (c, n = 5 mice for control; n = 4 mice for other groups), LW/BW (d, n = 5 mice for 378a-EVs; n = 4 mice for other groups) and serum triglyceride levels (e, n = 4 mice) were measured in young mice transplanted with macropahge-miR378a-EVs. f,g, GTT (f) and ITT (g) were performed on young mice treated with macropahge-miR378a-EVs (n = 9 mice). h, Outline of generating specific knockout of miR-378a in BMMs of miR-378aflox/flox mice (18 months old, male mice). i, The mRNA level of senescence-associated markers in muscle (n = 6 mice for AAV-Control; n = 5 mice for other groups) and adipose tissue (n = 5 mice). j, Protein levels of senescence markers in liver of miR-378a-BMM-CKO mice (left) and quantitative analysis (right) (n = 3 mice). k, GTT and ITT were performed on miR-378a-BMM-CKO mice (n = 6 mice for AAV-Control; n = 5 mice for other groups). The colored shading in f, g and k represents the area under the curve. l, Phosphorylation levels of insulin signaling in liver (left) and quantitative analysis (right) (n = 3 mice). m, Outline of the studies (22 months old, male mice). n, Representative SA-β-gal staining of femur from aged mice after infection with AAV-miRNA-191 (scale bar, 50 μm; n = 5 mice). The red arrows represent the SA-β-gal-positive cells. o,p, Representative µCT images (o) and quantitative analysis of bone volume/tissue volume (BV/TV) and cortical bone thickness (CT. Th) (p) (n = 12 mice). q, Outline of the studies (2 months old, male mice). r, Representative p53 fluorescence staining of femurs after infection with AAV-F4/80-miR-191-sponge (scale bar, 50 μm; n = 6 mice) (top) and quantitative analysis (right). s, Representative µCT images (left) and quantitative analysis (right) (n = 6 mice). Data are expressed as mean ± s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001 and #P < 0.0001, as determined by one-way ANOVA followed by Tukey’s multiple comparison test in c–g; two-tailed t-test in i (muscle and Cdkn1a, Il-6 and Cxcl1 in adipose), j–n, p (CT. Th) and s; and two-tailed t-test with Welch’s correction in i (Il-1β in adipose), p (BV/TV) and r. mo, months; TG, triglyceride; Rel. fold, relative fold; T.Ar, total area.
