Fig. 6: Supermeres affect the in vivo levels of liver lipids and glycogen.
a, Schematic of the mouse treatment experiments. D, day. b, Liver-to-body weight ratio of mice following PBS, exomere or supermere treatments. Two-sided Kruskal–Wallis test, followed by Dunn’s post-hoc test. Data are the mean ± s.e.m. of n = 6 animals. c, Oil red O staining of mouse livers following three consecutive injections with PBS (left) or exomeres (middle) and supermeres (right) derived from DiFi cells. The livers were harvested 24 h after the last injection. Scale bars, 20 µm. d, Level of triglycerides in liver tissue following injection with exomeres or supermeres derived from DiFi cells. e, Periodic acid–Schiff staining of formalin-fixed paraffin-embedded (FFPE) liver tissue following injection with exomeres or supermeres derived from DiFi cells. There were significant differences between experimental groups by pathology scoring of hepatocytes containing darker magenta deposits of polysaccharides (arrowheads; P = 0.038, two-sided Kruskal–Wallis test). Representative images are shown. Inset: magnified view with a diameter of approximately 90 µm. Scale bars, 100 µm. f, Histological scoring of liver sections stained with periodic acid–Schiff (PAS). The sections were scored double-blinded (0–3) for intensity and homogeneity by two liver pathologists. The liver sections from the mice injected with 300 µg of supermeres showed decreased scores in comparison to the other treatment groups. d,f, Two-sided Wilcoxon rank-sum test; n = 6 animals. For the boxplots, the centre lines mark the median, the box limits indicate the 25th and 75th percentiles, and the whiskers extend 1.5× the interquartile range from the 25th and 75th percentiles. g, Immunoblot of select proteins in mouse liver lysates after treatment with PBS (control) or 300 µg of exomeres or supermeres. h, Levels of proteins detected by immunoblot. Data are the mean ± s.e.m. of n = 3 animals. One-way ANOVA, followed by Holm–Bonferroni correction. i, Venn diagram of unique and common genes that are differentially expressed compared with the control (PBS) group between exomere- and supermere-treated mice. The criteria for inclusion of a differentially expressed gene were fold change > 1.5 and FDR < 1.0. j, Principal component (PC) analysis of gene expression in the mouse liver cells following treatment. Exom, exomere; super, supermere; CV, centrilobular vein; and CTL, control. *P < 0.05.
