Figure 5. Tnfr1 Signaling Promotes Premalignant Cholangiocellular Lesions.

(A) Cell proliferation of WT or Tnfr1^−/− cholangiocytes was analyzed by bromodeoxyuridine (BrdU) incorporation.

(B) Hepatoblasts were kept in basal medium or basal medium supplemented with DMSO or Tnf. IF of A6 and Hnf4α was performed. Scale bar, 20 μm.

(C) Survival analysis for Hspd1^ΔLPC, Hspd1^ΔLPC Tnfr1^+/−, and Hspd1^ΔLPC Tnfr1^−/− mice.

(D) Macroscopic appearance of 8-week-old Hspd1^ΔLPC, Hspd1^ΔLPC Tnfr1^+/−, and Hspd1^ΔLPC Tnfr1^−/− livers. Scale bar, 1 cm.

(E and F) H&E, A6 and Ki67 IHC, and Tnfr1 in situ hybridization of 8-week-old WT, Hspd1^ΔLPC, and Hspd1^ΔLPC Tnfr1^−/− livers (E), as well as quantification of necrosis and cholangiocellular overgrowth (F). The dashed lines indicate the border between bilary cells and hepatocytes. Scale bar, 50 μm.

(G) IHC of 8-week-old Hspd1^ΔLPC and Hspd1^ΔLPC Tnfr1^−/− livers. Scale bar, 20 μm.

(H) qRT-PCR of 6- and 8-week-old WT, Hspd1^ΔLPC, and Hspd1^ΔLPC Tnfr1^−/− livers for indicated genes.

(I) Percentage of Ki67⁺ hepatocytes in WT, Hspd1^ΔLPC, and Hspd1^ΔLPC Tnfr1^−/− livers.

Data are represented as the mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001. ns, not significant. See also Figure S5.