Fig. 6.

Juglans regia L. extract increased the expression of osteogenic genes and cell autophagy through the BMP2/Smad/Runx2 and Wnt/β-catenin pathways in hBMSCs. hBMSCs were treated with 2 mg/mL Dickkopf-1 (DKK1) or 100 ng/mL Noggin for 24 h and then treated with OIM (osteogenic induction medium, including 10 mmol/L β-glycerophosphate, 10 nmol/L dexamethasone, and 10 μg/mL ascorbic acid) and JRL extract (20 μM) for 7 days. The protein levels of BGLAP (a and b), osterin (a and c), OPG (a and d), LC3II (e and f), LC3I (e and f), Beclin-1 (e and g), and ATG5 (e and h) 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 < 0.05 vs OIM-cultured group, ^###P < 0.001 vs OIM-cultured group, ^&&P < 0.01 vs OIM + JRL extract-cultured group, ^&&&P < 0.001 vs OIM + JRL extract-cultured group