Fig. 2: Elevated extracellular Ca²⁺ induces the expression of FGF2, TGFβ1 and OPN in a time and dose-dependent manner.

a-c Time courses of FGF2, TGFβ1, and OPN expression after exposure to 8 mM Ca²⁺. C3H10T1/2 cells were incubated with standard medium (1.8 mM Ca²⁺) or 8 mM Ca²⁺ medium for the indicated time (a); 48 h (b); 72 h (c). d C3H10T1/2 cells were treated with a medium containing the indicated concentration of Ca²⁺ for 48 h. e BM-MSCs were incubated with standard medium or 6 mM Ca²⁺ medium for 48 h. a–e Cells were harvested and subjected to real-time RT-PCR to analyze expression levels of FGF2, TGFβ1, and OPN. Values for the relative expression of the indicated genes are expressed as the mean ± SEM of triplicate reactions in one representative experiment. a, c, and d Statistical analysis was performed by ANOVA followed by the Tukey post hoc test. **p < 0.01; ***p < 0.001 vs. first bar. ^##p < 0.01; ^###p < 0.001 vs. indicated group. b and e Statistical analysis was performed by Student’s t-test, *p < 0.05; **p < 0.01; ***p < 0.001 vs. first bar