Skip to main content
. 2023 Jun 2;30(9):1234–1248. doi: 10.1038/s41417-023-00633-y

Fig. 7. Clinical correlation between LBH protein expression and WNT hyperactivation in gastrointestinal cancers.

Fig. 7

Immunohistochemical (IHC) analysis of LBH protein expression (red/brown) in invasive colorectal carcinoma (CRC) specimen with validated anti-LBH antibody [9, 10]. A LBH is overexpressed with a predominant nuclear pattern in tumor cells at the invasive edge of CRC tumors (i–ii), compared to adjacent normal gut mucosa (iii). LBH-specific immunopositivity was also detected in tumor-associated stroma (i-ii). Gut-associated lymph follicles served as positive (+) control for LBH immunostaining (iv). B Representative IHC images of serial paraffin sections immunostained with LBH or ß-catenin antibodies. LBH expression in CRC cells at the invasive tumor front (Inv. Front) correlates with nuclear β-catenin staining, indicative of active WNT signaling. Note, that neither LBH nor β-catenin was expressed in the tumor center. C Western blot analysis of LBH, β-catenin, and TCF4 in CRC cell lines with no (RKO), low (HCT116), and high endogenous (Caco2, SW480) WNT activity, and in normal and tumorigenic pancreatic, esophageal, and gastric cell lines, as indicated. β-Actin (loading control). D Transcriptional luciferase reporter assays in the same cell lines above (C), using WNT responsive TOPFlash relative to FOPFlash reporter activity. Data are means ± SEM from three experiments performed in duplicates. P-values: *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001 (one-way ANOVA). E Western blot analysis of LBH protein expression in selected normal and tumorigenic GI cell lines above (C) 72 h after transient transfection of cells with β-catenin (CTNNB1) siRNA or scramble control (CTRL) siRNA.