Fig 2: CRBN modulates Wnt function in zebrafish.a A schematic of transverse views of a zebrafish head indicating phenotypes regulated by Wnt signaling. White arrows indicate eyes and the balbis indicates the distance between eyes. b The transverse views of a (i) WT zebrafish head, a WT zebrafish head injected with (ii) crbn mRNA, or (iii–v) one of three distinct crbn guide RNAs (sgRNA) along with dCas9 mRNA, in the absence or presence of (iv) crbn mRNA or (v) β-catenin (ctnnb1) mRNA. White arrows indicate eyes, the gray arrow indicates merged eyes, and the balbis indicates the distance between eyes. Scale bar = 200 μm. c, e and g Quantifications of the indicated eye phenotypes are shown. Asterisks indicate statistical significance (Fisher’s exact test, *p value < 0.05, ****p value < 0.0001). d and f RNA extracts from the indicated zebrafish (20 hpf) were used to determine the expression of three Wnt target genes using quantitative RT-PCR. Quantification of gene expression in indicated zebrafish embryos (mean ± SEM, n = 3 independent pools of zebrafish embryo) is shown. Asterisks indicate statistical significance (two-tailed Student’s t-test, *p value < 0.05, ***p value < 0.001, ****p value < 0.0001). lef1 lymphoid enhancer-binding factor 1.

Fig 3: Nuclear β-catenin expression is higher in the S45F mutated.a Expression of total β-catenin levels in a subset of desmoid tumor cell lines by western blot. b Expression of nuclear β-catenin levels in a subset of desmoid tumor cell lines by western blot. c Expression of total β-catenin levels in CTNNB1 transfected 293T cells by western blot. d Expression of nuclear β-catenin levels in CTNNB1 transfected 293T cells by western blot.

Fig 4: TCF7 knockdown neutralizes the intensity of ABC transporters and Wnt/ß-catenin signal in response to imatinib. (A) GSEA analysis shows the effects of TCF7 and imatinib on Wnt/ß-catenin signaling pathways in CML imatinib-resistant cells. (B) Heatmap of the genes in TCF7_KD core enriched ABC transporters and Wnt/ß-catenin signaling pathways. (C) Western blot analysis showing the expression of ABCC2, CCND1, CTNNB1 and TCF7 proteins. (D) RT-qPCR analysis showing the expression of ABCC2 and CCND1 mRNA in the Scramble and TCF7_KD groups. (E) Integrative genomics viewer (IGV) showing the recruitment of TCF7 to the ABCC2 promoter region and the transcription level of ABCC2 in K562/G01 cells with or without TCF7_KD treatment. (F) Recruitment of TCF7 in the ABCC2 promoter region shown by ChIP-qPCR. (G) Role of TCF7 and imatinib on the Wnt and ABC transporters signaling pathway in imatinib-resistant CML. One-way ANOVA with Tukey's post hoc test were performed for F. Two-tailed Student's t-test was used for I. *P<0.05, **P<0.01, ***P<0.001. ns, not significant. GSEA, Gene Set Enrichment Analysis; TCF7, transcription factor 7; CML, chronic myeloid leukemia; KD, knockdown; CCND1, cyclin D1; CTNNB1, catenin ß1; ACTB, ß-actin.

Fig 5: ß-catenin is frequently altered in HCC patient samples(B) Frequency of genetic alteration in four genes, CTNNB1, APC, AXIN1 and FGF19, were determined in 207 HCC patients from ASAN collection (A) and 373 HCC patients from TCGA collection (B). Red indicates presence of genetic alteration and gray indicates wild-type. (C) Representative immunohistochemical analysis of normal liver and HCC. (D) Boxed plot of ß-catenin cytoplasmic and nuclear staining (H-score) levels in human normal liver and HCC.