Fig. 6.

P-Tyr42 RhoA induces cell proliferation in response to Wnt3A. (A) HEK293T cells were stimulated Wnt3A (30 ng/ml) for 48 h. Cell were stained with DAPI (1 μg/ml) for 10 min, and DAPI fluorescence was measured with a fluorescence microscope. In addition, cell proliferation was also measured using MTT assay. (B, C) HEK293T and RAW264.7 cells, respectively, were pretreated with Tat-C3 (1 μg/ml) and Y27632 (10 μM) for 1 h and then treated with Wnt3A (30 ng/ml) for 2 days. Cell proliferation was measured by measuring DAPI fluorescence with a fluorescence microscope. In addition, cell proliferation was also measured using MTT assay. (D) HEK293T cells were transfected with RhoA WT, Y42E and Y42F (4 μg DNA) for 24 h, then treated with Wnt3A (30 ng/ml) for 2 days. MTT assay was used for cell proliferation. (E) HEK293T cells were pretreated with Tat-C3 (1 μg/ml) and Y27632 (10 μM) for 1 h and then treated with Wnt3A (30 ng/ml) for 2 h and cyclin D1 and c-Myc were detected by western blotting. (F) HEK293T cells were transfected with RhoA WT, Y42E and Y42F (4 μg DNA) for 24 h, then treated with Wnt3A (30 ng/ml) for 2 days and cyclin D1 and c-Myc were detected by western blotting.