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. Author manuscript; available in PMC: 2016 Jun 16.
Published in final edited form as: Cell Rep. 2015 Jun 4;11(10):1625–1637. doi: 10.1016/j.celrep.2015.05.019

Fig 3.

Fig 3

RANK-stimulated osteoclast formation requires ASXL2. A,B,C) ASXL2−/− BMMS, transduced with NFATc1 or vector, were exposed to M-CSF and RANKL for 5 days. WT BMMs transduced with vector served as control. The cells were A) stained for TRAP; B) osteoclasts counted; C) Osteoclastogenic proteins were determined. D) ASXL2−/− BMMS, transduced with NFATc1 or vector were exposed to M-CSF and RANKL for 1 day. NFATc1 and c-Fos expression was determined. E and F) Cytokine/serum starved BMMs were exposed to RANKL with time. E) Cytoplasmic osteoclastogenic signaling molecule activation and F) nuclear osteoclastogenic signaling molecule activation were determined by immunoblot. G) BMMs were maintained in M-CSF alone for 3 days (0) or M-CSF and RANKL for 48 or 72 hrs. RANK protein was measured. H) BMMs, transduced with hFas/RANK, were exposed to M-CSF and anti-Fas activating antibody for 5 days. The cells were stained for TRAP activity. I) c-Fos expression by WT and ASXL2−/− BMMs, transduced with hFas/RANK or vector and exposed to M-CSF and anti-Fas activating antibody, was determined. Scale Bar: 400 μm.