Enhanced Proliferation of Porcine Bone Marrow Mesenchymal Stem Cells Induced by Extracellular Calcium is Associated with the Activation of the Calcium-Sensing Receptor and ERK Signaling Pathway

Figure 2

Inhibition of CaSR reversed the promotive effects of on the proliferation of pBMSCs. (a) Effects of NPS2143 (0.1 μM), an antagonist of CaSR, on the proliferation of pBMSCs after 5-day incubation (). (b) Effects of 4 mM and/or 0.1 μM NPS2143 on the cell cycle progression of pBMSCs. After pBMSCs were exposed to 4 mM with or without NPS2143 (0.1 μM) for 5 days, the cells were collected and treated according to the protocol in Materials and Methods. The DNA contents were measured with FACScan flow cytometry. (c) Analysis of proliferation index (PI) and the percentage of cells in G0/G1, S, and G2/M phases. (d) The mRNA expression levels of cyclins (cyclin A2, cyclin D3, and cyclin E2), PCNA, and p21 in response to 4 mM and/or 0.1 μM NPS2143. (e) Western blot analysis of cyclin D1 and p21 in pBMSCs after 5-day culture. β-actin was used as loading control. (f) Mean ± SEM of immunoblotting bands of cyclin D1 and p21; the intensities of the bands were expressed as the arbitrary units (). , , and versus 1 mM group (control); , , and versus 4 mM group.

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