Fig. 1

The effect of different concentrations of Aubucin on viability and osteogenic differentiation in hBM-MSCs. a The chemical structure of Aubucin. b The proliferation of hBM-MSCs was measured by CCK-8 assay after cell cultured with different concentrations of Aubucin (0–1 μM) for 1, 3, 5 or 7 days in hBM-MSCs growth medium. c-h The relative mRNA expression of osteogenesis-related genes (COL1A1, RUNX2, OCN, ALP, SP7 and OPN) was measured by qRT-PCR on days 3 and 7 after osteogenic differentiation. i-k The relative protein expression of osteogenesis-related genes (COL1A1, RUNX2 and SP7) were evaluated by western blotting analysis after osteogenesis for 3 and 7 days and normalized to GAPDH. l, m, p Immunofluorescence staining for COL1A1 and RUNX2 proteins after 3 days of osteogenesis. Scale bars, 50 μm. n, o, q Different concentrations of Aubucin (0–1 μM) promoted hBM-MSCs mineralization and ALP activity. ALP staining and activity was detected on day 3 of osteogenesis. ARS staining and quantitation was detected on day 14 of osteogenesis. Scale bars, 500 μm. All of the experiments were performed in triplicates. Data are expressed as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 compared to the control group on day 3. ^#P < 0.05, ^##P < 0.01, ^###P < 0.001, ^####P < 0.0001 compared to the control group on day 7