Figure 6. JNK/c-Jun Activation Is Required for Premalignant Cholangiocellular Lesions.

(A) p-JNK, p-Stat3, and Rela IHC in 8-week-old WT and Hspd1^ΔLPC livers. The black arrowhead indicates a p-JNK⁺ cholangiocyte, the white arrowhead indicates a p-JNK⁺ hepatocyte. Black arrows indicate positive staining in non-parenchymal cells, while white arrows indicate negative staining in liver parenchymal cells. Scale bar, 20 μm.

(B) Rela and c-Jun IF in hepatoblasts cultured in basal medium supplemented with DMSO or Tnf. White arrowheads indicate c-Jun nuclear staining. Scale bar, 20 μm.

(C) Timeline of SP600125 administration on Hspd1^ΔLPC mice and survival of Hspd1^ΔLPC mice not treated (mock) or treated with SP600125.

(D) CK19 and Ki67 IHC in 8-week-old Hspd1^ΔLPC livers treated with mock or SP600125, and quantification of cholangiolar cancerous lesion areas and Ki67⁺ cholangiocytes relative to total cholangiocytes. Scale bar, 50 μm.

(E) c-Jun IF in hepatoblasts cultured in Tnf-containing medium with or without SP600125. White arrowheads indicate the c-Jun nuclear staining. Scale bar, 20 μm.

(F and G) Phase contrast and A6 and Hnf4α IF images (F) and qRT-PCR (G) of hepatoblasts in basal or Tnf-containing medium with or without SP600125. Scale bar, 20 μm.

Data are represented as the mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001. ns, not significant. See also Figure S6.