Skip to main content
[Preprint]. 2024 Nov 18:2024.11.18.624183. [Version 1] doi: 10.1101/2024.11.18.624183

Figure 2. CD73 restrains the transcriptional activity of Xenopus exon 3 mutant β-catenin.

Figure 2.

(A) NT5E mRNA and (B) protein (CD73) expression in EC cell line models. (C) Validation of TCF/LEF-luciferase reporter activity using TOPFlash/FOPFlash plasmids for endogenous β-catenin and Xenopus exon 3 mutant β-catenin (β-catenin∆EX3). (D) TCF/LEF reporter activity using CTNNB1 siRNA in HEC-1-A cells. (E) Immunoblot validation of siRNA knockdown of β-catenin in HEC-1-A cells. TCF/LEF reporter activity with or without Xenopus β-catenin∆EX3 transfection in (F) NT5E WT HEC-1-A cells and Ishikawa cells or (G) NT5E WT and NT5E KO HEC-1-A cells. (H) Immunofluorescence showing CD73 is localized to the membrane in Ishikawa cells overexpressing AdV-transduced NT5E. Membrane localization of CD73 in HEC-1-A cells serves as a positive control. Images are cropped from originals (originals are in Supplemental Figure 2) using BZ-X800 analyzer software (Keyence). Cropping was intended to more easily highlight membrane localization of CD73. All images were cropped to the same size from a 20X image. Scale bars 20 um. (I) TCF/LEF reporter activity in Ishikawa cells with or without Xenopus β-catenin∆EX3 transfection and AdV-transduced NT5E. (D, F, G, I) Graphs are pooled data from n = 3 independent experiments (D) or n = 2 independent experiments (F, G, I). Data represent the mean ± SEM. *P < 0.05, ****P < 0.0001; 2-way ANOVA with Sidak’s post test.