Fig. 5: Response of hepatic stellate cell (HSC)-specific Foxm1 knockout (Foxm1HSC−/−) mice to BDL.
a H&E, Sirius red, CK19, α-SMA, and F4/80 staining in Flox control and Foxm1HSC−/− mice after BDL as compared to sham surgery. Liver fibrosis was measured by Sirius red staining (n = 8 animals per group) (b) and hydroxyproline assay (n = 6 animals per group) (c), liver injury by ALT (n = 6 animals in Flox + Sham, Flox + BDL and Foxm1HSC−/− + BDL groups and n = 5 in Foxm1HSC−/− + Sham) (d) and AST (n = 7 in Flox + Sham, Flox + BDL groups and n = 6 in Flox + BDL group and n = 5 in foxm1HSC−/− + BDL) (e) levels, macrophage number by F4/80 (n = 4 animals per group) (f), and ductular proliferation by CK19 staining (n = 6 animals per group) and myofibroblast differentiation by α-SMA staining (n = 3 animals per group) (g). For b, Sirius red area/total area of Flox BDL vs. Sham and HSC−/− BDL vs. Flox BDL, p = 0.0000000015 and p = 0.0000060 respectively. For c, hydroxyproline (ug/g, liver) for Flox BDL vs. Sham and HSC−/− BDL vs. Flox BDL, p = 0.00000012 and p = 0.000094 respectively. For d, ALT level (ug/L) for Flox BDL vs. Sham and HSC−/− BDL vs. Flox BDL, p = 0.000000041 and p = 0.0000013 respectively. For e, AST level (ug/L) for Flox BDL vs. Sham and HSC−/− BDL vs. Flox BDL, p = 0.00000000066 and p = 0.00044 respectively. For f, F4/80 positive number for Flox BDL vs. Sham and HSC−/− BDL vs. Flox BDL, p = 0.0000000071 and p = 0.0073 respectively. For g, Flox BDL vs. Sham and HSC−/− BDL vs. Flox BDL for CK19/total area, p = 0.000000000075 and p = 0.080 respectively; for α-SMA area/total area, p = 0.000026 and p = 0.00011 respectively. Data are shown as mean ± SEM, ***p < 0.001, ****p < 0.0001. h Protein expression of FOXM1, MATα2, and MAT2β in hepatocytes, cholangiocytes, HSCs, and KCs isolated from Flox control and Foxm1HSC−/− mice ± BDL, Densitometry values for protein levels are summarized in Supplementary Fig. 11e, f. mRNA levels of Foxm1, Mat2a, and Mat2b in hepatocytes (n = 6 per group) (i), cholangiocytes (n = 6 per group) (j), HSCs (n = 6 per group) (k), and KCs (n = 6 per group) (l) isolated from Flox control and Foxm1HSC−/− mice ± BDL. Data are shown as mean fold of Flox control ± SEM, ****p < 0.0001, and ns not significant. p values obtained via two-tailed unpaired Student’s t tests. For i, mRNA levels in Hepatocytes, fold of Flox con of FOXM1, MAT2A, MAT2B for Flox BDL vs. Sham, p = 0.0000000000004, p = 0.0000000000001 and p = 0.0000000000072 respectively; for HSC−/− BDL vs Flox BDL, p = 0.000000000022, p = 0.000000000020, and p = 0.000000054 respectively. For j, mRNA levels in Cholangiocytes, fold of Flox con of FOXM1, MAT2A, and MAT2B for Flox BDL vs. Sham p = 0.0000000000000, p = 0.000000028, and p = 0.0000000085 respectively; for HSC−/− BDL vs Flox BDL, p = 0.17, p = 0.42, and p = 0.46 respectively. For k, mRNA levels in HSCs, fold of Flox con of FOXM1, MAT2A, and MAT2B for Flox BDL vs. Sham p = 0.0000052, p = 0.0000000000, and p = 0.0000000000 respectively; for HSC−/− BDL vs Flox BDL, p = 0.00000056, p = 0.00018, and p = 0.00000000002 respectively. For l, mRNA levels in KCs, fold of Flox con of FOXM1, MAT2A, and MAT2B for Flox BDL vs. Sham p = 0.000000000091, p = 0.00036, and p = 0.00000010 respectively; for HSC−/− BDL vs Flox BDL, p = 0.086, p = 0.61, and p = 0.15 respectively. Statistical significance was determined by using two-tailed unpaired Student’s t-test. Source data are provided as a Source Data file. Abbreviations: ALT alanine transaminase, AST aspartate aminotransferase, BDL bile duct ligation, Cho cholangiocytes, HSCs hepatic stellate cells; Hep hepatocytes, KCs Kupffer cells.
