Extended Data Fig. 4: Pepd silencing exacerbates metabolic disturbances in HFD 58%-fed mice.
Body weight curves and fat mass % curves of pepd WT, HET and KO mice after 20 weeks HFD 45% (n = 8, 11, 9 biologically independent animals per group, respectively) and HFD 58% (n = 8, 11, 8 biologically independent animals per group, respectively). b, c. Blood glucose levels up to 120 min. after an intraperitoneal injection of glucose in a glucose tolerance test (b) or insulin (0.75 IU/kg) in an insulin tolerance test (c) with the representative AUC in pepd WT (n = 9), HET (n-11) and KO (n = 9) mice fed HFD 45%. d. Fasting glucose, insulin and FFA blood levels in pepd WT (n = 8), HET (n = 11) and KO (n = 8) mice fed HFD 45%. e. Representative images of red Sirius staining in ScW and GnW from C57Bl/6 mice fed chow (n = 6), 45%HFD (n = 8) and HFD 58% (n = 8, 20 weeks) conditions and quantification of peri-adipocyte collagen content (peri-AD) represented in % Area (excluding perivascular staining).f. Heat map of gene expression in GnW from C57Bl/6 mice fed 20 weeks with chow (n = 6), 45%HFD (n = 8) and HFD 58% (n = 8). Results are expressed as fold change over chow diet. g. Representative images of blots and quantification of total and basal phosphorylated (Ser473) AKT in gastrocnemius muscle of pepd WT, HET and KO mice fed chow (n = 5, 6, 5 biologically independent animals per group, respectively), and HFD 58% conditions (n = 5, 5, 7 biologically independent animals per group, respectively). h. Heat map of gene expression of fibro-inflammatory and functional markers in gastrocnemius of pepd WT, HET and KO mice fed chow (n = 6, 7, 6 biologically independent animals per group, respectively), and HFD 58% conditions (n = 8, 7, 8 biologically independent animals per group, respectively). i. Heat map of gene expression in GnW from pepd WT, HET and KO fed HFD 45% (n = 6, 10, 9 biologically independent animals per group, respectively) and HFD 58% (n = 8, 9, 9 biologically independent animals per group, respectively) expressed as fold change variation over chow diet; *p < 0.05 compared to chow, #p < 0.05 compared to WT. j, k. Pathway enrichment analysis of the DEGs in GnW (i) and liver (j) from pepd KO mice (n = 9) compared to WT mice (n = 8) fed HFD 45%, using different data bases (KEGG, Reactome, Biocarta, NABA and PID). The heat maps indicate the level of significant changes (false discovery rate-adjusted p-value). Data is presented as mean values +/− SEM. Data was analysed using a 2-way ANOVA followed by a Turkey post-hoc multiple comparisons test; G, genotype; X, interaction (a-c, i), or a one way ANOVA followed by a Sidak (b, c, e) or Dunnett (d, g, h) post-hoc multiple comparisons test. A two-tailed Student’s t-test was also used to analyse the data (f).
