Wnt5a induces PI3K activation of osteosarcoma cells. (A) Human osteosarcoma cells MG-63, serum-deprived for 24 h, were untreated or treated with 100 ng/ml of Wnt5a and harvested at 15 min, 30 min, and 1 h after the start of treatment for SDS-PAGE and immunoblot analysis with antibodies to p-PI3K p85 (p-Tyr458) and total PI3K p85. p-PI3K/total PI3K ratios were indicated at each time. Data were presented as mean ± SD of 3 determinations. *, **: p < 0.05, p < 0.01 in the cultures with Wnt5a relative to the cultures without Wnt5a. (B) Serum-deprived MG-63 cells were pre-treated with 20 μM LY294002 for 1 h. Relative cell migration rate was determined by using wound healing assay in MG-63 cells incubated in the absence or presence of 100 ng/ml Wnt5a for 10 h. ** p < 0.01 in the cultures with Wnt5a relative to the cultures without Wnt5a. Data were presented as mean ± SD of 5 determinations.