Fig. 4
S1P2, S1P3, and Gα12/13 are required for Rho activation in astrocytes. WT astrocytes were transfected with an SRE.L luciferase reporter construct to assess Rho activation following knockdown with control siRNA or siRNA (2 μM) targeting S1P1, S1P2, S1P3, Gα12/13, or Gαq. Cells were also treated with pertussis toxin (PTX, 100 ng/mL, 24 h) to inhibit Gαi or with the C3 exoenzyme (0.5 μg/mL, 4 h) to inhibit Rho. Following knockdown or pretreatment, cells were stimulated with S1P (0.5 μM, 8 h). Control siRNA treated cells stimulated with S1P were set at 100% response. Data shown are mean ± SEM from four independent experiments run in duplicate. #p < 0.05 between control and siRNA S1P-treated groups