Skip to main content
. 2019 Feb 6;5:62. doi: 10.1038/s41420-018-0134-6

Fig. 2. Inhibition of Rankl induced osteoclastogenesis in macrophages from Nampt+/− mice.

Fig. 2

a Bone marrow-derived macrophages from Nampt+/+ and Nampt+/− mice were treated with M-CSF (20 ng/ml) for 2 days. Non-adherent cells were washed out and cells were further cultured with M-CSF (20 ng/ml) and Rankl (100 ng/ml) for an additional 3 days. TRAP staining was performed 3 days later and cells having 3 or more nuclei were counted as osteoclasts. The frequency of osteoclasts is expressed as mean ± SD (N = 5). An average of 7 fields/well were counted per experimental sample. b Reduced osteoclast target gene expression in Nampt-deficient macrophages from Nampt+/− mice. Osteoclast target Nfatc1, Acp5, Dc-stamp, Cathepsin K mRNA expression were measured using semi-quantitative RT-PCR. β-Actin was used as a loading control. c Representative western blot of Nampt protein in Nampt-deficient macrophages. Quantification of Nampt protein expression in Nampt-deficient macrophages. Representative images from three Nampt+/− mice with Nampt+/+ littermate controls are presented. Quantification of semi-quantitative RT-PCR of osteoclast target genes expressed as mean ± SD (N = 3). *P < 0.05; **P < 0.005