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. Author manuscript; available in PMC: 2022 Apr 1.
Published in final edited form as: J Hepatol. 2021 Dec 25;76(4):850–861. doi: 10.1016/j.jhep.2021.12.019

Fig. 4. BAs activate FXR in HSCs to promote liver fibrosis.

Fig. 4.

(A) Schematic representation of the Nr1h4HSC mouse model. (B) Relative mRNA levels of Nr1h4 of isolated HSCs from 14-week-old Nr1h4(+/+)HSC and Nr1h4(Δ/Δ)HSC mice (n=3). (C) WB of isolated HSCs from 14-week-old Nr1h4(+/+)HSC and Nr1h4(Δ/Δ)HSC mice. (D) Representative pictures of desmin/FXR co-IHC of livers from 14-week-old Nr1h4(+/+)HSC and Nr1h4(Δ/Δ)HSC mice. (E) Quantification of desmin/FXR co-IHC described in (D) (n=4,3). (F) Schematic representation of the bile duct ligation surgery protocol. (G) Representative pictures of Sirius Red staining of livers from 14-week-old Nr1h4(+/+)HSC and Nr1h4(Δ/Δ)HSC mice sham and bile duct-ligated (BDL). (H) Quantification of Sirius Red staining described in (G) (n=3,3,6,8). (I) Relative mRNA levels of indicated genes of isolated hepatocytes from Nr1h4(+/+)HSC mice sham and BDL (n=3). (J) WB of isolated hepatocytes from Nr1h4(+/+)HSC mice sham and BDL. (K) Schematic representation of the in vitro experiment performed in HepG2 and LX-2 cell lines. (L) Cholic acid levels measured in media from HepG2 cells transfected with siCtr or siMCRS1 and/or NTCP overexpression plasmid as indicated (N=4). (M) Relative mRNA levels of ACTA2 in LX-2 cells transfected with siCtr or siNR1H4 and treated with/without ChR, supplemented with conditioned medium from HepG2 cells transfected with siCtr or siMCRS1 and pDONR-Ctr or pDONR-NTCP as indicated (N=3). (N) Relative mRNA levels of COL1A1 in LX-2 cells transfected with siCtr or siNR1H4 and treated with/without ChR, supplemented with conditioned medium from HepG2 cells transfected with siCtr or siMCRS1 and pDONR-Ctr or pDONR-NTCP as indicated (N=3). (O) Cholic acid levels measured in media from isolated hepatocytes from Mcrs1hep mice and Mcrs1(Δ/Δ)hep mice overexpressing NTCP (N=3). (P) Relative mRNA levels of Acta2 in isolated HSCs from Nr1h4HSC mice supplemented with conditioned medium from isolated hepatocytes from Mcrs1hep mice and Mcrs1(Δ/Δ)hep mice overexpressing NTCP (N=3). (Q) Relative mRNA levels of Col1a1 in isolated HSCs from Nr1h4HSC mice supplemented with conditioned medium from isolated hepatocytes from Mcrs1hep mice and Mcrs1(Δ/Δ)hep mice overexpressing NTCP (N=3). Scale bar: 50 μm in (D) and 100 μm in (G). Statistical analysis was performed using unpaired two-tailed Student’s t test in (B) and (I), and two-way ANOVA with Tukey’s multiple comparisons test in (H), and (L) to (Q). Data are represented as means±SEM. *, p≤0.05; **, p≤0.01; ***, p≤0.001.