Fig. 5

Effect of EECM on BMP-related pathways during osteoblastogenesis. a Western blot analysis of MAPK signaling, WNT, and BMP2/4. The effect of the BMP antagonist, Noggin, on (b) ALP activity, and (c) western blot analysis of osteoblast differentiation markers in MC3T3-E1 cells pretreated with noggin (200 ng/mL) for 1 h, followed by treatment with EECM and retreated after 3 days. d Effect of EECM on ERE activity in MCF7 human breast adenocarcinoma cells. Cells were transfected with the ERE and CMV plasmid DNAs and analyzed for luciferase activity after EECM treatment. The effect of the ER antagonist, ICI182.780, on (e) ALP activity and (f) western blot analysis of osteoblast differentiated markers in MC3T3-E1 cells pretreated with ICI182.780 (10 μM) for 1 h, followed by treatment with EECM and treated after 3 days. *p < 0.05, **p < 0.01, ***p < 0.001, compared to differentiated cells