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
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.
5), suggesting p38 pathway plays a negative role in HBMSCs cultures.
RSVL-induced ER signaling and MAPK activations result in osteogenic genes expression in HBMSCs cultures.