Fig. 4.

Wnt5a stage-dependently utilizes the NFAT and NF-κB pathways in a stage-dependent manner. A and B, Wnt5a stage-dependently regulates NFAT and NF-κB nuclear localization in a stage-dependent manner. C17 micromass cultures were treated with Wnt5a or vehicle control (Ctl) as indicated and cultured for 21 d. Cultures were harvested and fractionated into nuclear and cytoplasmic extracts. Western blotting for levels of NFAT, NF-κB, and CREB in each fraction was performed (A), and the nuclear to cytoplasmic ratio was determined (B) Values are the mean of three independent experiments ± sem. *, P < 0.05. C and D, Wnt5a differentially regulates NFAT and NF-κB Activity. C17 cells were transfected with NFAT (C), NF-κB (D), or vector control luciferase (LUC) reporter constructs and treated with Wnt5a as indicated. After each treatment, cells were harvested in passive lysis buffer, and luciferase activity was determined. *, P < 0.05.