Expression of hDvl3, but neither hDvl1 nor hDvl2, rescues Wnt5a-stimulated NF-AT-sensitive transcriptional activation in Dvl3-depleted cells. F9 cells stably expressing Fz2 were treated with siRNA targeting Dvl3. Dvl3-depleted cells were transfected with various amounts of expression plasmid harboring hDvl3-eGFP (A and B), or hDvl1-eGFP or hDvl2-eGFP (C and D). The Wnt5a-sensitive transcriptional activation was performed by the assay of NF-AT-dependent luciferase reporter (A and C). ***, p ≤ 0.001, compared with basal (-Wnt5a) in control (-siRNA, -plasmid) group; ##, p ≤ 0.01, ###p ≤ 0.001, compared with (+Wnt5a) in control group. The cell lysates from the same batch used in reporter assay were subjected to SDS-PAGE and the resolved proteins transferred to nitrocellulose blots. The blots were stained with antibodies indicated (B and D). A set of representative images of immunoblotting are shown.
Ma et al. Journal of Molecular Signaling 2010 5:19 doi:10.1186/1750-2187-5-19