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. 2017 Jun 3;8(34):55967–55983. doi: 10.18632/oncotarget.18356

Figure 3. Melatonin decreased NAMPT-dependent Sirt1 expression in IL-1β-insulted chondrocytes.

Figure 3

A. Mock- or IL-1β-treated chondrocytes were pretreated with 100 nM siNAMPT and incubated in serum-free media overnight. NAMPT protein expression was measured by Western blot. β-actin was used as internal control. B.-E. Chondrocytes were pretreated with 100 μM NAMPT inhibitor (FK866) for 30 min or 100 nM siNAMPT for 1 h and then stimulated with or without 10 ng/ml IL-1β for 30 min, followed by 10 ng/ml melatonin for 24 h. (B) mRNA and (C) protein levels of Sirt1 were measured by qPCR and Western blot assays, respectively. GAPDH and β-actin were used as internal controls. D. Quantification of Sirt1 protein expression in (C). E. Sirt1 activity was determined. Each value represents means ± SD of 3 replicates or representative of 3 independent experiments. *P < 0.05 compared with control; #P < 0.05 compared with IL-1β-treated group; §P < 0.05 compared with Siscrb group. Mel: melatonin.