Fig. 6: Response of Kupffer cell (KC)-specific Foxm1 knockout (Foxm1^KC−/−) mice to BDL.

a H&E, Sirius red, CK19, α-SMA, and F4/80 staining in Flox control and Foxm1^KC−/− 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 = 5 animals in Flox + Sham group and n = 6 in foxm1^KC−/− + Sham, Flox + BDL and foxm1^KC−/− + BDL groups) (c), liver injury by ALT (n = 6 animals per group) (d) and AST (n = 6 animals per group) (e) levels, macrophage number by F4/80 (n = 6 animals per group) (f), and ductular proliferation by CK19 (n = 4 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 KC−/− BDL vs. Flox BDL, p = 0.00000000049 and p = 0.00014 respectively. For c, hydroxyproline (ug/g, liver) for Flox BDL vs. Sham and KC−/− BDL vs. Flox BDL, p = 0.00030 and p = 0.012 respectively. For d, ALT level (ug/L) for Flox BDL vs. Sham and KC−/− BDL vs. Flox BDL, p = 0.0000000000000 and p = 0.000000000081 respectively. For e, AST level (ug/L) for Flox BDL vs. Sham and KC−/− BDL vs. Flox BDL, p = 0.0000000000001 and p = 0.00000025 respectively. For f, F4/80 positive number for Flox BDL vs. Sham and KC−/− BDL vs. Flox BDL, p = 0.0000000067 and p = 0.0000094 respectively. For g, Flox BDL vs. Sham and KC−/− BDL vs. Flox BDL for CK19/total area, p = 0.00000010 and p = 0.039 respectively; for α-SMA area/total area, p = 0.000026 and p = 0.00033 respectively. Data are shown as mean ± SEM, *p < 0.05, ***p < 0.001, ****p < 0.0001. h Protein levels of FOXM1, MATα2, and MAT2β in hepatocytes, cholangiocytes, HSCs, and KCs isolated from Flox and Foxm1^KC−/− mice ± BDL, Densitometry values for protein levels are summarized in Supplementary Fig. 12a–d. 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 Foxm1^KC−/− mice ± BDL. Data are shown as mean fold of Flox control ± SEM, ***p < 0.001, ****p < 0.0001, ns not significant. For i, mRNA levels in hepatocytes, fold of Flox con of FOXM1, MAT2A, MAT2B for Flox BDL vs. Sham, p = 0.000000012, p = 0.0000019 and p = 0.50 respectively; for KC^−/− BDL vs Flox BDL, p = 0.00000086, p = 0.00060, and p = 0.0000000036 respectively. For j, mRNA levels in cholangiocytes, fold of Flox con of FOXM1, MAT2A, and MAT2B for Flox BDL vs. Sham p = 0.0000000000001, p = 0.0000000000005, and p = 0.00000000046 respectively; for KC^−/− BDL vs Flox BDL, p = 0.071, p = 0.064, and p = 0.061 respectively. For k, mRNA levels in HSCs, fold of Flox con of FOXM1, MAT2A, and MAT2B for Flox BDL vs. Sham p = 0.0000000086, p = 0.0000000002, and p = 0.000000000 respectively; for KC^−/− BDL vs Flox BDL, p = 0.0000032, p = 0.0000095, and p = 0.0000078 respectively. For l, mRNA levels in KCs, fold of Flox con of FOXM1, MAT2A, and MAT2B for Flox BDL vs. Sham p = 0.000000040, p = 0.000000033, and p = 0.00000075 respectively; for KC^−/− BDL vs Flox BDL, p = 0.00000000071, p = 0.00000097, and p = 0.0000059, 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.