Wnt3a can further enhance the activation of β-catenin in Twist-overexpressing cells. (a) Expression of β-catenin before or after Wnt3a stimulation (Wnt3a conditioned medium for overnight) was measured by Western blotting. β-tubulin served as a loading control. (b) The activation of β-catenin before and after Wnt3a stimulation (Wnt3a-conditioned medium for overnight) was examined by measuring the transcription activity of Top/Fop luciferase in Hela cells and their corresponding stable transfectants (mean ± SD of three separate experiments). *P < 0.05 for Hela cells compared with the corresponding Twist-overexpressing cells. #
P < 0.01 for Twist/Hela cells compared with cells treated with Wnt3a. Non-significant (ns) was found between Hela cells treat with and without Wnt3a. (c) CD44 promoter luciferase plasmid was expressed in Hela cells and their corresponding Twist-overexpressing stable transfectants. After 36 hr, cells were treated overnight with a Wnt3a-conditioned medium and luciferase activity was measured by using a Dual-Luciferase Reporter Assay (Promega) (mean ± SD of three separate experiments). * and # P < 0.05. Non-significant (ns) was found between Hela cells treat with and without Wnt3a.