Involvement of PI3K in SWT extract-induced bone mineralization in osteoblasts. (A) Cells were seeded in 24-well plates and cultured for 2 wks in medium containing vitamin C (50 μg/mL) and β-glycerophosphate (10 mM). The cells were concomitantly treated with SWT extract (150 μg/mL) plus Ly294002 or wortmannin. Mineralized nodule formation was assessed by alizarin red-S staining. (B) Cells were incubated with SWT extract (150 μg/mL) plus Ly294002 or wortmannin for 72 h, and ALP was measured with an ALP activity assay kit. (C) Cells were pretreated with Ly294002 and wortmannin for 30 min or transfected with p85 siRNA for 24 h followed by stimulation with SWT extract for 24 h, and ALP, BMP-2, and OPN mRNA expression was measured by qPCR. (D) Cells were treated with SWT extract for the indicated time intervals, and p-p85 phosphorylation was examined by western blotting. *, p < 0.05 compared to the control group; #, p < 0.05 compared to the SWT extract-treated group.