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Fig. 4

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KLEXJ promotes ALP activity and BMP-2 expression through the NF-κB pathway. (A) Osteoblasts were pretreated with TPCK (10 μM) or PDTC (10 μM) for 30 min, followed by stimulation with KLEXJ for 48 h. ALP activity was examined using a commercial ALP assay kit. (B-D) Osteoblasts were pretreated with TPCK (10 μM) or PDTC (10 μM) for 30 min, followed by stimulation with KLEXJ for 24 h and BMP-2 expression was examined by qPCR and Western blot analysis. (E) Osteoblasts were incubated with KLEXJ for indicated time intervals and p65 phosphorylation was examined by Western blot analysis. (F) Osteoblasts were pretreated with Ly294002 or an Akt inhibitor, followed by stimulation with KLEXJ. p65 phosphorylation was examined by Western blot analysis. Results are expressed as mean ± S.E.M. *, P < 0. 05 compared with control. #, p < 0.05 compared with KLEXJ-treated group.