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. Author manuscript; available in PMC: 2015 Dec 1.
Published in final edited form as: J Bone Miner Res. 2014 Dec;29(12):2618–2635. doi: 10.1002/jbmr.2295

Fig. 6. Comparison of the effects of TNF-α and RANKL on osteoclastogenesis.

Fig. 6

(A–D) Quantitative PCR of osteoclast-associated genes. BMMs were stimulated with TNF-α (100 ng/ml) and RANKL (10 and 50 ng/ml). Gene expression levels of Acp5 (A), Ctsk (B), Oscar (C), and Nfatc1/A (D) were determined. Gene expression levels relative to Hprt were calculated and normalized to the expression level of Sh3bp2+/+ BMMs at 0 hour. (E, F) Resorption assay. BMMs were cultured with TNF-α or RANKL at indicated concentrations for 14 days on dentin slices. After removal of the cells, resorption areas were visualized by toluidine blue (E). Resorbed areas per total surface area were quantified (F). (G) Western blot analysis for NFATc1. BMMs were stimulated with TNF-α (100 ng/ml) or RANKL (10 and 50 ng/ml) for 72 hours. NFATc1 expression levels in nuclear and cytoplasmic fractions were determined. Data are presented as mean ± SD. +/+: Sh3bp2+/+, KI/+: Sh3bp2KI/+. # P < 0.05 compared to TNF-α-stimulated Sh3bp2KI/+ BMMs at each time point.