Research Paper Volume 13, Issue 4 pp 5150—5163

Nampt promotes osteogenic differentiation and lipopolysaccharide-induced interleukin-6 secretion in osteoblastic MC3T3-E1 cells

Nampt promoted LPS-induced inflammation in MC3T3-E1 cells. After treatment for 3 days, (A) Real-time PCR analysis showing the mRNA expression of IL-6, Nampt and Sirt1 normalized to β-actin. (B) ELISA result showing the IL-6 levels. (C) NAD+/NADH analysis showing intracellular content of NAD+ and NADH, and their ratio. (D) Western blot analysis showing the protein expression of Nampt, Sirt1, TAK1 and p-NF-κB p65. (E) The nuclear translocation of NF-κB p65 was measured by fluorescence immunocytochemistry (200×). Red, NF-κB p65; blue, DAPI. The data are expressed as the mean ± SD. *, pppp

Figure 3. Nampt promoted LPS-induced inflammation in MC3T3-E1 cells. After treatment for 3 days, (A) Real-time PCR analysis showing the mRNA expression of IL-6, Nampt and Sirt1 normalized to β-actin. (B) ELISA result showing the IL-6 levels. (C) NAD+/NADH analysis showing intracellular content of NAD+ and NADH, and their ratio. (D) Western blot analysis showing the protein expression of Nampt, Sirt1, TAK1 and p-NF-κB p65. (E) The nuclear translocation of NF-κB p65 was measured by fluorescence immunocytochemistry (200×). Red, NF-κB p65; blue, DAPI. The data are expressed as the mean ± SD. *, p<0.05; **, p<0.01. #, p<0.05 vs. con; ##, p<0.01 vs. con.