Fig. 2

Juglans regia L. extract concentration-dependently promoted osteoblastic differentiation of hBMSCs. hBMSCs were treated with OIM (osteogenic induction medium, including 10 mmol/L β-glycerophosphate, 10 nmol/L dexamethasone, and 10 μg/mL ascorbicacid) and different concentrations of Juglans regia L extract (10, 20, 40, or 80 μM) for 1–14 days. a The cell viability was determined using CCK-8 solution for the cells treated by different concentration of JRL extract (10, 20, 40, or 80 μM). b Alkaline phosphatase (ALP) activity of hBMSCs was detected by a commercial ALP assay kit. c hBMSCs were treated with different concentrations of JRL, fixed and stained with Alizarin red. d–g The protein levels of Osteocalcin (BGLAP), Osterix, and osteoprotegerin (OPG) were tested by Western blot analysis. β-actin is a loading control. Data were represented as mean ± SD from at least three independent experiments performed for each assay. **P < 0.01 vs control group, ***P < 0.001 vs control group