FANCL overexpression enhances β-catenin activity and protein expression. (A) Flow cytometry analysis of LEF-TCF-eGFP reporter assays in 293FT cells transfected with vector-control, LacZ-control, or human FANCL cDNA. Cells were treated with BIO (a GSK3-β inhibitor that activates the Wnt pathway). Data shown are from 4 experiments (mean ± SEM), *P < .01; **P < .05 compared with vector-control and LacZ-control. (B) Immunoblot analysis to quantify β-catenin in cells transfected with FANCL or vector-control. Shown are representative immunoblots for the cytoplasmic (left panel) and nuclear (right panel) fractions and a graph displaying the quantitation from 7 independent blots (mean relative expression to the no BIO and no FANCL control ± SEM), *P = .004; **P = .03; ***not significant; ****P = .008. Loading controls and the purity of the subcellular fractions were evaluated by staining for α-tubulin (n = 6) and Sp1 (n = 5) levels. Shown are representative blots at similar exposure times. We also performed statistical analysis to show that there were no differences in these controls for the pairwise comparisons performed (P values for student t test analysis of tubulin and Sp1 levels were between .77 and .98) and there were no differences globally among the 4 experimental conditions tested (P values for ANOVA analysis of tubulin and Sp1 levels were .36 and .49, respectively).