Figure 3.

Cyclic mechanical loading could regulate osteogenic differentiation and NOTCH signaling in human BMSCs. (a) qRT-PCR analysis and (b) western blotting analysis of osteogenic differentiation markers ALP, COL1a1, and OCN in BMSCs after treatment with 10% CMS for 3 weeks compared with static control cells. GAPDH was used as an internal control. (c) Representative images of ALP staining (including quantitative analysis) and (d) Alizarin red staining of BMSCs after treatment with 10% CMS for 3 weeks compared with static control cells. (e) Immunostaining of NICD (green) and RUNX2 (red) location in BMSCs after treatment with 10% CMS for 3 weeks compared with static control cells. Scale bar, 50 μm. (f) qRT-PCR analysis of JAG1 expression in BMSCs after treatment with 10% CMS for 3 weeks compared with static control cells. (g) qRT-PCR analysis of NOTCH1~4, JAG1, JAG1, HES1, and HEY1 mRNA levels and (h) western blot analysis of BMSCs after treatment with 10% CMS for 3 weeks compared with static control cells. (i) qRT-PCR analysis and western blotting analysis of HDAC1 in BMSCs after treatment with 10% CMS for 3 weeks compared with static control cells. All results are representative of at least three independent experiments. All the staining data were confirmed by three repeated tests. Data were mean±S.D., *P<0.01, **P<0.001. All P-values are based on Student's t-test