Fig. 4

S1P pre-conditioning boosts intracellular calcium in hypoxia-impaired C2C12 cell differentiation. S1P (1 μM) and VC pretreated differentiating C2C12 cells under normoxia (N) and 0.5% hypoxia (H) were stained with Fluo-4 dye at pre-defined study time points. Intracellular calcium was quantified by a fluorimeter. Data are reported as relative fluorescence signal intensity with respect to 0D, which is proportional to the calcium binding to Fluo-4. Data are reported as mean ± SD from three independent experiments carried out in triplicate. 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