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

Fig. 3. Gpr126 regulates osteoblast proliferationdifferentiation, and mineralization.

Fig. 3

(A and B) Bone marrow stromal cell (BMSC) proliferation and differentiation were inhibited in Gpr126-deficient osteoblasts as determined by colony-forming unit (CFU) assay. BMSCs from 1-month-old Osx-Cre;Gpr126fl/fl mice and Ctrl littermates were cultured for 14 days and then subjected to crystal violet staining (CFU-F, left) or Alizarin red staining (CFU-Ob, right). Representative images are shown (A). The number (n) of colonies per well for CFU-F and CFU-Ob was counted (B). Scale bars, 10 mm. *P < 0.05. n = 3. (C) BMSC differentiation and mineralization were suppressed in Gpr126 deletion osteoblasts. BMSCs were isolated from 1-month-old Osx-Cre;Gpr126fl/fl mice and Ctrl littermates and subjected to ALP staining (7 day), von Kossa staining (14th day), and Alizarin red staining (21st day) assays. Scale bars, 5 mm. (D) Gpr126 KO inhibited ALP enzyme activity (n = 5) and Ocn relative mRNA expression (n = 2) in osteoblasts. BMSCs were isolated from 1-month-old Osx-Cre;Gpr126fl/fl mice and Ctrl littermates and differentiated into osteoblasts. The cells were harvested at days 7 and 14 of differentiation for ALP enzyme activity assay and Ocn mRNA quantitation by real-time PCR, respectively. *P < 0.05, ***P < 0.001. n = 5.