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. 2010 Sep 24;285(48):37427–37435. doi: 10.1074/jbc.M110.149484

FIGURE 1.

FIGURE 1.

TNF can neither induce osteoclastogenesis nor activate osteoclast genes. A, BMMs were treated with M-CSF (M, 44 ng/ml), M-CSF (44 ng/ml) and RANKL (100 ng/ml) (M+R), or M-CSF (44 ng/ml) and TNF (10 ng/ml) (M+T) in a tissue culture dish for 4 days (d). The cultures were then stained for TRAP activity. B, BMMs on bone slices were cultured with M-CSF (44 ng/ml), M-CSF (44 ng/ml) and RANKL (100 ng/ml), or M-CSF (44 ng/ml) and TNF (10 ng/ml) for 4 days, and bone slices were then stained with Hoechst 33258 (Hoechst) or Alexa Fluor 488 phalloidin (Phalloidin). A separate set of cultures was continued for 4 additional days to perform bone resorption assays. C, BMMs were treated with M-CSF (44 ng/ml), M-CSF (44 ng/ml) and RANKL (100 ng/ml), or M-CSF (44 ng/ml) and TNF (10 ng/ml) in a tissue culture dish for 1, 2, or 4 days. Gene expression was determined by semiquantitative RT-PCR.