Figure 3.

TNFα modulates S1P signalling. Quantitative mRNA analysis was performed by real-time PCR in total RNA extracted from C2C12 myotubes stimulated or not with 100 ng/mL TNFα for the indicated time intervals (6 h and 24 h). mRNA quantitation of S1PR (S1P₁, S1P₂, S1P₃, S1P₄ and S1P₅) and S1P specific transporter, Spns2, was based on the 2^−ΔΔCt method, using individual enzyme or Spns2 of the unchallenged specimen as calibrator. Data are the mean ± SEM of three independent experiments performed in triplicate. The increase in S1P₂ and S1P₃ and the decrease in Spns2 expression induced by TNFα were statistically significant by Student’s t test (* p < 0.05; ** p < 0.01).