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. 2024 Aug 19;43(21):4846–4869. doi: 10.1038/s44318-024-00196-0

Figure 2. BATokine PCPE-1 promotes liver fibrosis in MASH.

Figure 2

Littermate control (Con) or BAT-specific Pcolce knockout mice (BAT Pcolce KO) were fed a high-fat diet (HFD) from 4 weeks of age and studied at 42–43 weeks of age. (A) Quantitative PCR (qPCR) results showing transcript Pcolce in BAT from indicated mice (n = 6, 6). (B) Western blot analysis for procollagen C-endopeptidase enhancer-1 (PCPE-1) in BAT from indicated mice. The quantification results for this panel see Fig. EV2C. (C) ELISA for plasma PCPE-1 (n = 6, 6) in the indicated mice aged 42–43 weeks. (D) Western blot analysis for PCPE-1 in liver from indicated mice. The right panel indicates the quantification (n = 3,5). (E) Picrosirius red staining of liver from the indicated mice. Scale bar = 500 μm. For the quantification results see Fig. EV2G. (F–H) ELISA for liver collagen type I (F) (n = 5, 5), liver triglyceride (TG) (G) (n = 35, 27), and plasma alanine transaminase (ALT) (H) (n = 8, 8) in the indicated mice aged 42–43 weeks. (I) Results from qPCR showing transcripts Col1a1 (n = 6, 6), Ccn2(Ctgf) (n = 5, 6), Tgfb1 (n = 5, 6), Ccl2 (n = 5, 6), and Tnf (n = 6, 6) in liver from indicated mice. (J) Experimental design of the BAT-adeno-associated virus (AAV) mouse: Pcolce gain-of-function model (AAV-mPcolce). AAV-mPcolce was injected into BAT at 50 and 54 weeks of age (a total of 2 injections) in a normal chow (NC) fed mice and studied at 58 weeks of age. (K) Results of a qPCR showing transcript Pcolce in BAT (n = 5, 5). (L, M) Western blot analysis for PCPE-1 in BAT (n = 5, 5) (right panel indicates the quantification of PCPE-1 relative to Tubulin) (L), and ELISA for mouse PCPE-1 in plasma (M) (n = 5, 5) of indicated mice. (N) Picrosirius red staining of liver from the indicated mice. Scale bar = 500 μm. For the quantification results see Fig. EV2J. (O–Q) ELISA for liver collagen type I (O) (n = 5, 5), liver TG (P) (n = 5, 5), and plasma ALT (Q) (n = 5, 5) from indicated mice. (R) qPCR of Col1a1 (n = 5, 5), Ccn2(Ctgf) (n = 4, 5), Tgfb1 (n = 5, 5), Ccl2 (n = 4, 5) and Tnf (n = 5, 5) in liver from the indicated mice. All data were analyzed by an independent-samples T-test. Data information: Representative data of two or more independent series (A, D, F–H), one independent series (I, K, L, R), one independent experiment analyzing samples from at least 2 independently prepared samples (C, F–H, M, O–Q). *p < 0.05, **p < 0.01. Values represent the mean ± SEM. NS = not significant. All data are from different biological replicates. Source data are available online for this figure.