Extended Data Fig. 7: Purified PEPD protein promotes fibro-inflammation in pre-adipocytes and stellate cells through EGFR signaling.

a. Representative images of confocal analysis of anti-Collagen type I (in red,) and Bodipy staining (lipid accumulation in green) (i) and corresponding quantifications of collagen I staining (j) in 3T3-L1 adipocytes (n = 4 biologically independent samples) pre-treated (Erlo) or not (PBS) with Erlotinib (5 µM) prior treatment with (PEPD) or without (control) purified PEPD protein (250 nM) during the first 5 days of adipogenic differentiation. * compared to control PBS, # compared to PEPD PBS. X, interaction. b. Il-6 level in culture media of mature adipocytes isolated from ScW of C57Bl/6 mice treated or not (control) with purified PEPD protein (250 nM, 24 h, n = 6 biologically independent animals per group). c. Representative images of confocal analysis of anti-Collagen type IV (in green,) and Actin staining (in red) and corresponding quantifications of collagen IV staining in stellate cells (n = 3 biologically independent samples) pre-treated (Erlo) or not (PBS) with Erlotinib (5 µM) prior treatment with (PEPD) or without (control) purified PEPD protein (250 nM). d. Gene expression profile in stellate cells (n = 4) pre-treated (Erlo) or not (PBS) with Erlotinib (5 µM) prior treatment with (PEPD) or without (control) purified PEPD protein (250 nM). e. AST serum level in PEPD-injected mice compared to controls (n = 11/group). f. Representative images of H&E and Sirius staining in the liver from PEPD-injected mice compaired to control (n = 11 biologically independent animals per group, a), and quantification of lipid droplet (steatosis) and collagen contents (fibrosis). g. Liver gene expression profile in PEPD-injected mice compared to controls (n = 11 biologically independent animals per group). h. Representative images of red Sirius staining in gastrocnemius from PEPD-injected mice compaired to control (n = 11 biologically independent animals per group), and quantification of collagen content (fibrosis). i. Gastrocnemius gene expression profile in PEPD-injected mice compared to controls (n = 11 biologically independent animals per group). j. Gene expression profile in muscle fibroblasts pre-treated (Erlo) or not (PBS) with Erlotinib (5 µM) prior treatment with (PEPD) or without (control) purified PEPD protein (250 nM, 5 days). k. Representative images of confocal analysis of anti-Collagen type I (in green,) and αSMA staining (in red), and corresponding quantifications of collagen I and αSMA stainings in muscle fibroblasts (n = 4 biologically independent samples) pre-treated (Erlo) or not (PBS) with Erlotinib (5 µM) prior treatment with (PEPD) or without (control) purified PEPD protein (250 nM, 5 days). Data is presented as mean values +/− SEM. Data was analysed using a 2-way ANOVA followed by aTukey post-hoc multiple comparisons test (a, c, d, j, k), or using a two-tailed Student’s t-test (b, e-i).