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. 2020 Mar 20;6(12):eaaz0368. doi: 10.1126/sciadv.aaz0368

Fig. 5. COLIV is an activating ligand of Gpr126 to regulate osteoblast differentiation and mineralization.

Fig. 5

(A) COLIV stimulated cAMP production in Ctrl osteoblast but not in Gpr126 deletion osteoblasts. COLIV (1 μM) was coated on 24-well plates, and then the BMSCs were seeded for differentiation. After 14 days, the cells were harvested and subjected to cAMP ELISA assay. **P < 0.01, ***P < 0.001. ns, no significant difference. n = 3. (B) COLIV-induced ALP enzyme activity (n = 3) and OCN mRNA expression (n = 2) in Ctrl osteoblasts but not in Gpr126 deletion osteoblasts. ns, not significant; *P < 0.05; ***P < 0.001. (C) COLIV-stimulated osteoblast differentiation and mineralization in Osx-Cre Ctrl (Ctrl) osteoblasts but not in Gpr126 deletion osteoblasts. ALP staining, von Kossa staining, and Alizarin red staining of BMSCs were performed after 7, 14, and 21 days of differentiation, respectively, while treated with or without COLIV. Scale bars, 5 mm.