Extended Data Fig. 5: Pepd silencing in hematopoietic cells prevent obesity and associated metabolic disturbances.
a. Prolidase activity in BMDMs during differentiation (n = 4 biologically independent samples). b. Pepd mRNA relative expression in adipocytes (AD), Mɸ (CD11b positive cells) and negative stroma-vascular fraction (SVF−) isolated from GnW of WT (n = 6) and LepOb/Ob mice (n = 9). *compared to AD WT, # compared to Mφ WT. c. Abundance of PEPD measured by mass spectrometry in unstimulated human iPS-derived Mɸ differentiated from FPS10C iPS line. Relative abundance of PEPD in comparison of other detected protein in the same sample is plotted in the graph- where actin and cyclin are representing examples of high and low abundant proteins respectively. d, e. Pepd mRNA expression (d) or PEPD ELISA in culture media from BMDMs treated or not (M0) 6 h (d) or 24 (e) with LPS, dexamethasone (GC) or IL4 (n = 4 biologically independent samples). BMDMs were treated with LPS for 1,6 or 24 h: f, g. PEPD level in culture media (f) and prolidase activity in BMDMs (g). h. Prolidase activity in culture media from BMDMs treated without (control) or with LPS (100 ng/ml, 24 h) (n = 4 biologically independent samples). I, j. Blood glucose levels up to 120 min. after an intraperitoneal injection of glucose (2 g/kg) in a glucose tolerance test (i) or insulin (0.75 IU/kg) in an insulin tolerance test (j) in BMT WT (n = 8) and KO mice (n = 6) fed chow. Respective AUC are represented. k-n. Fasting glucose (k), fasting insulin (l), fasting FFA (m) and fed glucose (n) blood levels in BMT-WT (n = 8) mice compared to BMTKO mice (n = 6) fed chow and HFD 58% (20 weeks).. o, p. Tissue weight (o) and Fat mass % (p) in BMT + / + (n = 8) and −/− (n = 6) mice fed chow and HFD 58% for 20 weeks. q. Body weight curve in BMT-WT (n = 8) mice compared to BMTKO mice (n = 6) mice between 0 and 20 weeks HFD 58%. r. Representative images of red Sirius staining ScW and GnW from BMT-WT (n = 8) mice compared to BMTKO mice (n = 6) fed HFD 58% and quantification of peri-AD collagen represented as %. s. Representative images of blots and quantification of total and basal phosphorylated (Ser473) AKT in GnW of of BMT WT and KO mice fed HFD 58% (n = 6/group). Data is presented as mean values +/− SEM. Data was analysed using a One way ANOVA followed by a Dunnett (a, f, g) or Tuckey (d, e) post-hoc multiple comparisons test, or using a 2-way ANOVA followed by a Tukey (b, k—p, r) or Sidak (I, j, q) post-hoc multiple comparisons test; G, genotype; X, interaction. A two-tailed Student’s t-test was also used to analyse the data (h-j, s).
