Fig 2: Expression of PCOLCE and PCOLCE2 mRNA and PCPE-1 protein in human liver samples.PCOLCE (A) and PCOLCE2 (B) mRNA expression in liver of control (n = 8) and NASH (F3-F4) patients (n = 5). PCPE-1 protein (C) in liver of control, NASH (F3-F4) and HCV cirrhotic patients (n = 4/group). One representative blot is shown, and the quantification was done from 3 independent experiments. Values reported from Western blots were obtained by band density analysis using Image Lab software (Bio-Rad) and expressed as the ratio of PCPE-1 / GAPDH. Data are expressed as mean ± SEM. **p<0.01 vs. control. Unpaired t test.

Fig 3: Liver steatosis and inflammation in WT and Pcolce-/- male mice under A04 or CDA-HFD after 8 weeks.A: Typical examples of liver histology with Hematoxylin and Eosin staining for inflammation and steatosis analysis (bars represent 100 µm except Pcolce-/- A04: 75 µm); B: evaluation of NAS score; C: steatosis score and D: inflammation score. Results are expressed as percentage of frequencies (C & D). NAS, steatosis and inflammation scores were determined as described in Material and Methods (n = 10–18). §§§p<0.001 vs. WT A04; °°°p<0.001 vs. Pcolce-/- A04. One-way ANOVA followed by Tukey’s post test for B. Fisher exact test for C & D.

Fig 4: Liver mRNA expression of lipogenesis, inflammation and fibrosis genes in male mice under A04 or CDA HFD after 8 weeks.Gene expression by RT-qPCR analysis in WT and Pcolce-/- male mice (n = 9–16). A: Fatty acid/ cholesterol synthesis (Acetyl-CoA carboxylase 1 (Acaca), Fatty Acid Synthase (Fsn), Sterol regulatory element-binding protein 1 (Srebf1)). B: Inflammation (CD68, Interleukin 1β (Il1b), Tumor Necrosis Factor α (Tnf)). C: Fibrosis (Actin α2 (Acta2), procollagen type I α1 (Col1a1), Lysyl Oxidase-like-2 (Loxl2)). Data are expressed as mean ± SEM. §§§p<0.001 vs. WT A04; °°°p<0.001 vs. Pcolce-/- A04; **p<0.01 vs. WT CDA HFD. One-way ANOVA with Tukey’s post-test.

Fig 5: Body and liver weight, liver TG, ALT and AST levels in WT and Pcolce-/- male mice under A04 or CDA HFD after 8 weeks.A: Body weight gain during A04 or CDA HFD diet (n = 12–18). §§§p<0.001 WT A04 vs. WT CDA HFD; °°°p<0.001 Pcolce-/- A04 vs. Pcolce-/- CDA HFD; *p<0.05 WT A04 vs. Pcolce-/- A04, Two-way ANOVA with Tukey’s post-hoc analysis. B: Liver weight (n = 12–18); C: Liver triglyceride (TG) content (n = 11–16); D: Plasma Alanine Aminotransferase (ALT) level (n = 8–15); E: Plasma Aspartate Aminotransferase (AST) level (n = 8–15). Panels B-E: §§§p<0.001 vs. WT A04; °°°p<0.001 vs. Pcolce-/- A04; *p<0.05 vs. WT CDA HFD. One-way ANOVA with Tukey’s post hoc analysis. Data are expressed as mean ± SEM.