Figure 10.

Wnt family members differentially regulate β-catenin phosphorylation and subcellular localization in C2C12 cells. (A-D) C2C12 cells were cultured with or without K252a, and then phospho-β-catenin (Y654) was immunostained. K252a caused an accumulation of vesicles containing phospho-β-catenin (Y654) in C2C12 cells. (E-L) Wnt3a, Wnt4 and Wnt5a are overexpressed in C2C12 cells by adenovirus-mediated gene transfer. After 2 day culture in differentiation medium commencing 24 h post-transfection, subcellular phospho-β-catenin (Y654) localization was determined by immunostaining. (M-T) C2C12 cells expressing eGFP, Wnt3a, Wnt4 and Wnt5a were immunostained with anti-pan-β-catenin antibodies. Cytoplasmic β-catenin is elevated with Wnt3a expression in proliferation medium and maintained at a higher level in differentiation medium, whereas nuclear and membrane-bound β-catenin is increased with Wnt4 expression.