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. 2019 Jul 31;33(11):12124–12134. doi: 10.1096/fj.201900733RR

Figure 1.

Figure 1

Osteoblast differentiation induced by BMPs. A) Matrix mineralization of MC3T3-E1 cells and mouse primary osteoblasts stimulated with or without BMP2, BMP4, or BMP9. Cells were induced to differentiate with or without 50 ng/ml BMP2, BMP4, or BMP9 in combination with 10 mM β-glycerophosphate for the indicated days. Cells were fixed and stained with 1% Alizarin Red S as described in Materials and Methods. B) Transcriptional activation of ALP, Hey1, Runx2, and Smad7 by BMP2, BMP4, and BMP9. After serum starvation for 4 h, MC3T3-E1 cells were stimulated with BMP2, BMP4, or BMP9 (50 ng/ml) for the indicated hours. Total RNA was isolated and reverse transcribed. Quantitative real-time PCR analyses were performed in triplicate using the specific primers for ALP (Alpl), Hey1, Runx2, and Smad7. Rpl13a was used as the endogenous control for normalization. Fold increase represents an experimental value divided by the 0 h control (untreated) value. Data represent means ± sd of triplicate samples. Each experiment was repeated at least 3 times with similar results. *P < 0.05 (significant difference from the same hour value of the BMP9 stimulation group by Mann-Whitney U test; n = 6).