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

Figure 7.

Figure 7

Essential roles of endoglin and GIPC1 in Akt-GSK3-β signaling pathway and ALP and Runx2 mRNA induction by BMP9. MC3T3-E1 cells were transfected with control siRNA, endoglin siRNA, or GIPC1 siRNA, followed by 48 h incubation at 37°C. A) Total RNA was isolated, reverse transcribed, and analyzed by real-time PCR analyses for endoglin and GIPC1 mRNA expressions. Relative mRNA expression levels were calculated based on the GAPDH mRNA level. Fold induction in comparison with the control siRNA-transfected cell mRNA is shown. Data represent means ± sd. *P < 0.05 (significant difference by Mann-Whitney U test; n = 6). B) Cells were serum starved for 4 h, followed by stimulation with 50 ng/ml BMP9 for the indicated minutes. Western blotting analyses were performed using the indicated antibodies. C) Cells were serum starved for 4 h and stimulated with 50 ng/ml BMP9 for the indicated hours. Total RNAs were isolated, reverse transcribed, and analyzed by real-time PCR analyses. Relative ALP and Runx2 mRNA expression levels were calculated based on the Rpl13a mRNA level. Fold mRNA induction in comparison with unstimulated control is shown. P, phosphorylated. Data represent means ± sd. *P < 0.05 [significant difference from the same day value of the control (Cont) siRNA group by Mann-Whitney U test; n = 6].