Figure 2. CaSR transcript relative abundance in eUCM-MSCs upon Ca²⁺- or calcimimetic-induced CaSR stimulation.

Quantitative Real Time RT-PCR analysis of the CaSR transcript in Ca²⁺ (2.5 mM CaCl₂) and calcimimetic-treated (2.5+R-467) equine UCM-MSCs versus controls (CTRL) or cells pre-incubated with the CaSR antagonist (2.5+2390+R-467). Cells were examined on day 4^th after treatments. For each sample, data (mean ± SD of three independent experiments in duplicate, CaSR average Ct) were normalized relatively to the abundance of HPRT1 mRNA (endogenous control) and normalized values were compared among groups. In the large cell line (panel A), CaSR transcription was strongly down-regulated in presence of additional Ca²⁺ (2% of control value; vs b: P<0.001) and NPS R-467 (13%; a vs b: P<0.001). In the small cell line (panel B), CaSR transcription was again down-regulated by additional Ca²⁺ (25%; a vs b: P<0.001) and NPS R-467 (72%; a vs b: P<0.001) even at a lesser extent. In both cell lines, the CaSR antagonist NPS 2390 reversed the effects of NPS R-467 (38% and 97%, for the large and the small cell line, respectively; c vs d: P<0.05).