Kaempferol-stimulated osteogenic effect is not mediated by activation of the Wnt/β catenin pathway. A: Cultured osteoblasts were treated with kaempferol (10 μM) for 2 days, with or without DKK-1 pre-treatment (0.2 mg/mL) for 1 hour. Total mRNA was extracted from the cultures to perform quantitative PCR for osteogenesis-associated genes, including type I collagen (COL1A1), osteonectin, osteocalcin, Runx2 and osterix. B: A reporter construct corresponding to pWRE-Luc (upper panel) was used as described previously. pWRE-Luc was transfected into cultured osteoblasts for 2 days before addition of Wnt3a (200 ng/mL; the ligand of Wnt/β-catenin pathway) or kaempferol (10 μM). Forty-eight hours later, luciferase activity was assayed. Values are expressed as the fold increase from the basal reading (control culture; 0.02% DMSO); mean ± SD, n = 5, each with triplicate samples.