Nox overexpression activates canonical Wnt signaling, but does not promote differentiation.

(A) Dual-luciferase assay of TCF activity in F9 cells transfected with EV and treated with DMSO or RA, or transfected with pcDNA3.1-Nox1 or pcDNA3.1-Nox4. (B) Dab2 expression in F9 cells transfected with EV and treated with DMSO or RA, or transfected with pcDNA3.1-Nox1. (C) Dab2 expression in F9 cells transfected with EV and treated with DMSO or RA, or transfected with pcDNA3.1-Nox4. (D) Immunoblot analysis for DAB2, TROMA1, and OCT4 in F9 cells transfected with EV and treated with DMSO or RA, or transfected with pcDNA3.1-Nox1, pcDNA3.1-Nox4 or both vectors. β-actin was used as a loading control. A total of 3 independent experiments were analyzed and results are presented as mean ± SEM. Letters denote groups of significance (P < 0.05) tested by a One-Way ANOVA followed by a Tukey’s test.