Figure 3. Sirt6 positively regulated osteoclastogenesis.

(a) BMMs from Sirt6^fl/flMx1Cre mice were transduced with pMX-puro (control, EV) or Flag-tagged Sirt6 retrovirus by stimulation with RANKL (50 ng/ml) for 3 days in the presence of M-CSF. Protein lysates were subjected to immunoblot analysis with Sirt6 and NFATc1 antibody. (b) Transduced BMMs were stained for TRAP after 5 days. Scale bar, 200 μm. (c) Number of TRAP⁺ MNCs (>5 nuclei) was counted as osteoclasts. (d) As in (a), except that BMMs from Sirt6^fl/fl mice were used. Protein lysates were subjected to as in (a). (e) Transduced BMMs were stained as in (b). Scale bar, 200 μm. (f) Number of TRAP⁺ MNCs (>5 nuclei) was counted as in (c). GAPDH was used as a loading control. *P < 0.05. Data are represented as mean ± S.D.