Fig. 3

Morusin promotes the Wnt/β-catenin signaling pathway in BMSCs. a Detection of the expression of target genes in Wnt/β-catenin pathway via quantitative real-time PCR. We normalized the gene expression levels to GAPDH. b, c DKK-1 partially reversed the pro-expression effect of Morusin (10 μM) on Col1a1, Runx2, and β-catenin. The protein expression level is the gray value ratio of the target protein to GAPDH. Gene expression was normalized to GAPDH. d ALP staining of the osteogenic BMSCs. BMSCs were treated with Morusin (10 μM) and DKK-1(500 ng /ml) for 3 days, fixed and stained with ALP staining. Scale bar, 250 μm. e Alizarin red staining of the osteogenic BMSCs. The cells were incubated with Morusin (10 μM) and DKK-1(500 ng/ml) for 14 days, and the calcium deposits were recognized via Alizarin red staining. Scale bar, 250 μm. f BMSCs immunofluorescence results of RUNX2 and COL1A1. Scale bars, 100 μm. BMSCs, bone marrow mesenchymal stem cells; RUNX2, runt-related transcription factor 2; COL1A1, collagen type I alpha 1; Dvl-1, dishevelled-1; LEF-1, lymphoid enhancing factor-1; NC, negative control; DKK-1, Dickkopf-related protein-1. We repeated the independent experiment for three times. Means ± SEM. *P < 0.05 and **P < 0.01 in comparison to the control group. “ns” refers to no statistical significance in comparison to the control group