Fig. 5
S1P3 and Sphk1 are upregulated in response to in vitro scratch injury and mediate COX-2 expression. In WT astrocytes, a S1P1, S1P2, S1P3 and b SphK1 mRNA expression were measured 1 h after scratch injury on culture plates. Data shown are mean ± SEM from three independent experiments run in triplicate. Fold increase is relative to the scratch control for each receptor subtype or Spkh1. *p < 0.05 between control and scratch-treated groups. COX-2 protein levels were measured after 8 h of scratch in c S1P3 KO astrocytes or d WT astrocytes pretreated with SPM-354 (5 μM, 15 min). COX-2 was normalized to GAPDH and expressed relative to the WT or KO controls or the ±inhibitor controls. Data shown are mean ± SEM from three independent experiments run in triplicate. e COX-2 protein levels were measured in WT astrocytes after knockdown of Sphk1 with siRNA (2 μM) or control siRNA followed by scratch for 8 h. COX-2 was normalized to GAPDH and expressed relative to the ±siRNA controls. Data shown are mean ± SEM from three independent experiments run in duplicate. * p < 0.05 and **p < 0.01 between control and scratch-treated groups and #p < 0.05 and ##p < 0.01 between scratch-treated groups of either WT and KO, with or without inhibitor, or siRNA