H]]-thymidine incorporation, ALP activity and calcium deposition assays were performed in HBMSCs culture as described above (Song et al.
Total RNA was isolated from the HBMSCs by the single-step method using Trizol reagent for quantitative real-time RT-PCR as described previously (Pan et al.
Effects of RSVL on cell proliferation and osteogenic differentiation in HBMSCs cultures
The peak stimulation of ALP activity was found at day 12 in osteogenic differentiation medium, both E2 and RSVL treatment significantly enhanced ALP activity as a function of time in HBMSCs cultures (data not shown).
1), indicating that ER mediated RSVL-induced anabolic responses in HBMSCs cultures.
Effects of RSVL on the activation of ERK1/2 and p38 in HBMSCs cultures
6] M) alone quickly activated both ERK1/2 and p38 activity in 5min in HBMSCs cultures, which lasted about 15min, then returned to basal levels (Fig.
5C) in HBMSCs cultures compared to SB203580 alone groups, but there were no further inductions in ALP activity (Fig.
Effects of RSVL on mRNA expression of RUNX2, Osterix and Osteocalcin in HBMSCs
Effects of RSVL on the expressions of RUNX2/CBFA1, Osterix and Osteocalcin were evaluated by real-time RT-PCR analysis in RNA preparations from HBMSCs cultures at day 8.
RSVL-stimulated proliferation and osteoblastic differentiation was via ER signaling pathway in HBMSCs cultures.
RSVL-induced ER signaling couples to MAPK pathways in HBMSCs cultures.