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Fig. 4 | Stem Cell Research & Therapy

Fig. 4

From: Aucubin promotes bone-fracture healing via the dual effects of anti-oxidative damage and enhancing osteoblastogenesis of hBM-MSCs

Fig. 4

Aubucin reverses oxidative damage and osteogenesis inhibition caused by exposure to 300 μM H2O2. a, b The relative protein expression of osteogenesis-related genes (COL1A1 and RUNX2), components of different signaling pathways (Total NRF2, HO1 and NQO1) and apoptosis-related genes (Bcl2, BAX and CC3) were evaluated by western blotting analysis after osteogenesis for 3 days and normalized to β-actin. c, d Representative micrographs and quantification of DCFH-DA staining of control group, H2O2 group, and H2O2 + AU (1 μM) group. Scale bars, 100 μm. e–h Expression of COL1A1 and RUNX2 proteins was determined by immunofluorescence on day 3 of osteogenesis. Scale bars, 50 μm. i, j Nuclear Nrf2 and cytoplasmic Nrf2 were quantified in hBM-MSCs after 3 days of treatment with AU and/or H2O2 by western blot analysis and normalized to THOC1 and β-actin. k, n Expression of Nrf2 protein was determined by immunofluorescence on day 3 of osteogenesis. Scale bars, 100 μm. l, m, o Aubucin (1 μM) promoted hBM-MSCs mineralization and ALP activity exposed to 300 μM H2O2. 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 H2O2 group. #P < 0.05, ##P < 0.01, ###P < 0.001 compared to the control (NC) group

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