Fig. 4: Response of hepatocyte specific Foxm1 knockout (Foxm1^Hep−/−) mice to BDL.

a H&E, Sirius red, CK19, α-SMA, and F4/80 staining in Flox control and Foxm1^Hep−/− mice after BDL as compared to sham surgery. Liver fibrosis was measured by Sirius red staining (n = 7 per group) (b) and hydroxyproline assay (n = 5 per group) (c), liver injury by ALT (n = 6 per group) (d) and AST (n = 6 per group) (e) levels, macrophage number by F4/80 (n = 6 per group) (f), and ductular proliferation by CK19 (n = 4 per group) and myofibroblast differentiation by α-SMA staining (n = 3 per group) (g). For b, Sirus red area/total area for Flox BDL vs. Sham and Hep−/− BDL vs. Flox BDL, p = 0.00000000011 and p = 0.0000015, respectively. For c, hydroxyproline (ug/g, liver) for Flox BDL vs. Sham and Hep−/− BDL vs. Flox BDL, p = 0.000058 and p = 0.0034, respectively. For d, ALT level (ug/L) for Flox BDL vs. Sham and Hep−/− BDL vs. Flox BDL, p = 0.000000000014 and p = 0.00000000097, respectively. For e, AST level (ug/L) for Flox BDL vs. Sham and Hep−/− BDL vs. Flox BDL, p = 0.0000000000001 and p = 0.00000025, respectively. For f, F4/80 positive number for Flox BDL vs. Sham and Hep−/− BDL vs. Flox BDL, p = 0.0000000020 and p = 0.00080, respectively. For g, CK19/total area for Flox BDL vs. Sham and Hep−/− BDL vs. Flox BDL, p = 0.00000078 and p = 0.00026, respectively. Data are shown as mean ± SEM, **p < 0.01, ***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 Foxm1^Hep−/− mice ± BDL, Densitometry values for protein levels are summarized in Supplementary Fig. 11a–d. mRNA levels of Foxm1, Mat2a, and Mat2b in hepatocytes (n = 6 animals in Flox + Sham, Foxm1^hep−/− + Sham, Flox + BDL groups and Foxm1^hep−/− + BDL group for foxm1 and mat2b mRNA; n = 4 in Flox + Sham, Foxm1^hep−/− + Sham, Flox + BDL groups and n = 6 in foxm1^hep−/−+BDL group for mat2a mRNA) (i), cholangiocytes (n = 4 animals in Flox + Sham, foxm1^hep−/− + Sham, Flox + BDL groups and foxm1hep−/− + BDL groups for foxm1 mRNA; n = 5 in Flox + Sham and foxm1^hep−/− + Sham groups and n = 4 in Flox + BDL and Foxm1^hep-−/− + BDL groups for mat2a mRNA; n = 6 in Flox + Sham, Foxm1^hep−/− + Sham, Flox + BDL groups and foxm1^hep−/− + BDL group for mat2b mRNA) (j), HSCs (n = 6 animals) (k), and KCs (n = 4 animals in Flox + Sham, Flox + BDL groups, foxm1^hep−/− + BDL groups and n = 5 in foxm1^hep−/− + Sham for foxm1 mRNA; n = 5 in Flox + Sham and foxm1^hep−/− + Sham groups, Flox + BDL and Foxm1^hep−/− + BDL groups for mat2a mRNA; n = 5 in Flox + Sham, Flox + BDL and Foxm1^hep−/− + BDL groups and n = 6 in foxm1^hep−/− + Sham group for mat2b mRNA). l isolated from Flox control, Foxm1^Hep−/− mice ± BDL. Data are shown as mean fold of Flox control ± SEM, *p < 0.05, ***p < 0.001, ****p < 0.0001. p values obtained via two-tailed unpaired Student’s t tests. For i, mRNA levels in hepatocytes, fold of Flox con of FOXM1, MAT2A, and MAT2B of Flox BDL vs. Sham p = 0.0000045, p = 0.000030, and p = 0.000028 respectively; of Hep^−/− BDL vs. Flox BDL p = 0.0000000099, p = 0.00024, p = 0.000099 respectively. For j, mRNA levels in cholangiocytes, fold of Flox con of FOXM1, MAT2A, and MAT2B of Flox BDL vs. Sham p = 0.00023, p = 0.00000026, and p = 0.000000011, respectively; of Hep^−/− BDL vs. Flox BDL p = 0.000041, p = 0.013, and p = 0.0000031, respectively. For k, mRNA levels in HSCs, fold of Flox con of FOXM1, MAT2A, and MAT2B of Flox BDL vs. Sham, p = 0.0000000000005, p = 0.000000067, and p = 0.0000000000 respectively; of Hep−/− BDL vs. Flox BDL p = 0.00000000050, p = 0.00025, and p = 0.0000000011 respectively. For l, mRNA levels in KCs, fold of Flox con of FOXM1, MAT2A, and MAT2B of Flox BDL vs. Sham, p = 0.000041, p = 0.000010, and p = 0.00000035 respectively; of Hep−/− BDL vs. Flox BDL p = 0.84, p = 0.16, and p = 0.62 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.