FOX transcription factors regulate Wnt pathway agonist expression.A, heat map of WNT and R-spondin gene expression changes following FOX gain-of-function. Gene expression was determined in 293T cells pretreated with 5 ng/ml recombinant human R-spondin 3. Each cell represents the average of three biological replicates, normalized to empty vector (EV) control. B and C, TOPflash assay in 293T following FOXP (B) or FOXN (C) overexpression. Where indicated, cells were treated with 10 nM LGK974 to block WNT protein secretion. D and E, grouping of TOPflash (D) and qPCR results (E) from Figure 1 by clusters of Wnt pathway agonist expression. Data in panel (A) were analyzed using Dunnett’s post hoc test against EV following one-way ANOVA (∗∗∗p < 0.001, ∗∗p < 0.01, ∗p < 0.05). RLA, relative luciferase activity. See also Fig. S4.