Fig. 7
Hypoxia modulates S1P receptor expression in C2C12 differentiating cells. S1P (1 μM) and VC pretreated differentiating C2C12 cells under normoxia (N) and 0.5% hypoxia (H) were analyzed for S1P receptor expression. Quantitative mRNA analysis was performed by using real-time PCR of the target genes S1PR1 (a), S1PR2 (b), S1PR3 (c) and S1PR4 (d). S1PR5 expression was not detected. Relative mRNA expression of the respective receptor was calculated after normalization to an endogenous reference gene (β-actin). Results are reported as fold changes (mean ± SD) from three independent experiments carried out in triplicate with respect to undifferentiated myoblasts (0D) at each time point according to the comparative CT method (\(2^{{ - \Delta \Delta C_{\text{T}} }}\) method). Statistical analysis was performed using the one-way ANOVA/post hoc Bonferroni’s analysis method. Statistical significance is denoted by NVC vs. HVC ($p < 0.05, *p ≤ 0.005) and HVC vs. HS1P (@p < 0.05, #p ≤ 0.005) at respective time points