Fig. 4.

Stat3 activation by HDL via S1P2 and S1P3 in DU145 cells. A: mRNA levels of S1P1, S1P2 and S1P3 were evaluated by real time PCR and expressed as RQV compared to Cyc. Values are expressed as the mean +SD (n=3). B; Protein levels of S1P2 and S1P3 were evaluated by western blotting. C–D: DU145 cells were preincubated in the absence (−) or presence of JTE013 (5 μM) for 1 h, and VPC23019 (5 μM) for 1 h. Cells were incubated with or without HDL (500 μg/ml) for 15 min, and whole cell lysate was analyzed by western blotting (Panel B). The graph shows the ratio of Stat3-ser-P to Stat3 in each sample relative to 1.0 for lane 2 (n=3, mean +SD). *P<0.05 vs lane 2. (Panel C). Cells were wounded and then cultured for 0 or 48 h with or without HDL (500 μg/mL). Values are expressed as the mean +SD *P<0.05 vs HDL (Panel D). After culturing DU145 cells with HDL, the numbers of cells that invaded through the matrigel were microscopically counted. Values are expressed as the mean +SD (n=3). *P<0.01 vs HDL (Panel E).