Extended Data Fig. 9: MAPK activity modulates liver zonation by promoting zone-1 and suppressing zone-3, inhibiting MAPK signalling in established tumours switches liver zonation resulting in a wnt high state and reduced mTOR activity.
From: Hepatic zonation determines tumorigenic potential of mutant β-catenin
a, Schematic representing liver-specific mouse model of acute Wnt (Rnf43fl/fl;Znrf3fl/fl), and BrafV600E activation. b, Representative images of BrdU and GLUL IHC of day-10 livers, n = 6. c, LW/BW ratios. d-g, BrdU, GLUL, CDH1, and SOX9 histo-scoring of day-10 livers. Bars are mean ± s.d. One-way ANOVA with Tukey’s multiple comparisons test. Biological replicates: (c) WT n = 9, RZ n = 10, Br n = 10, BrRZ n = 13; (d) WT n = 9, RZ n = 10, Br n = 9, BrRZ n = 10; (e) WT n = 7, RZ n = 8, Br n = 6, BrRZ n = 9; (f) WT n = 9, RZ n = 8, Br n = 10, BrRZ n = 11; (g) WT n = 8, RZ n = 8, Br n = 10, BrRZ n = 11. h, Schematic representing model of acute Wnt and BrafV600E activation. i-k, quantification of BrdU and GLUL in hepatocytes 4 days after recombination of Rnf43fl/fl;Znrf3fl/fl, BrafV600E or a combination of these alleles. Bars are mean ± s.d. Biological replicates: WT n = 10, RZ n = 8, Br n = 7, BrHETRZ n = 7 and BrHomRZ n = 8. (i, j) One-way ANOVA with Tukey’s multiple comparisons test. (k) two-sided, paired, t-test. l-n, quantification of BrdU and GLUL expression in hepatocytes 4 days after recombination of Ctnnb1ex3/WT, BrafV600E or a combination of these alleles. Bars are mean ± s.d. Biological replicates: WT n = 10, B n = 15, Br n = 7, BrHETB n = 10 and BrHomB n = 3. (l, m) One-way ANOVA with Tukey’s multiple comparisons test. (n) Two-sided, paired, t-test. WT and Ctnnb1ex3/WT data (i, j, l, m) also plotted in Fig. 3c and Extended Data Fig. 4c,e. WT and BrafV600E data repeated in figure panels i, j, l and m. o, Model of short-term dabrafenib treatment in established BrRZ tumours. The illustrations of the mouse and adenovirus in panels a,h,o were adapted from Medical Art Servier (https://servier.com) under a CC BY 4.0 licence. p, BrdU, GLUL and CDH1 IHC. q, Liver-to-body weight ratios. Biological replicates: day 60 n = 4; Vehicle, n = 11; dabrafenib, n = 10. r, Macroscopic tumour scoring. Biological replicates: day 60 n = 4; Vehicle, day 74 n = 8; dabrafenib, day 74 n = 7. Bars are mean ± s.d. One-way ANOVA and Dunn’s multiple comparisons test. s-t, SOX9, pS6(Ser235/236), pS6(Ser240/244) and p4E-BP1 (Thr37/46) IHC. Dashed line, highlights tumour boundary. Males, blue points; Females, red points. All scale bars, 100 μm. The illustrations of the mouse and adenovirus were adapted from Medical Art Servier (https://servier.com) under a CC BY 4.0 licence.
