Figure 6. Osteoclast differentiation induced by forced expression of RANK and TRAF6 in BMMs under adherent and non-adherent condition.

BMMs over-expressing RANK (transduced with pMX-RANK-puro vector) or TRAF6 (transduced with pMX-TRAF6-puro vector) were cultured with M-CSF (50 ng/ml), TGF-β (1 ng/ml), and RANKL (150 ng/ml) for 4 days under the adherent or non-adherent condition, then fixed and stained for TRAP. Cell under non-adherent condition were harvested and placed on cell culture plates for 1 hour to stain for TRAP.