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. Author manuscript; available in PMC: 2018 Nov 18.
Published in final edited form as: Bone. 2017 Oct 12;107:104–114. doi: 10.1016/j.bone.2017.05.009

Fig. 5.

Fig. 5.

C/EBPα or PU.1 overexpression fails to mediate osteoclast differentiation with the mutational inactivation of the RANK cytoplasmic 535IVVY538 motif. A, a schematic of the experimental strategy. B and C, gene expression analysis from BMMs doubly expressing FAS-RANK or FAS-mIVVY plus a GFP control (GFP), FLAG-C/EBPα (C/EBPα), or FLAG-PU.1 (PU.1) cultured with M-CSF for 4 days by RT-PCR (B) and Western blotting (C). D, TRAP staining for osteoclast differentiation from BMMs doubly expressing FAS-RANK or FAS-mIVVY plus GFP, C/EBPα, or PU.1 treated with M-CSF and α-FAS for 4 days. Scale bar = 200 μm. E , quantifications for D are shown. F and G, quantifications of the osteoclast size for D via the number of nuclei (F) and area (G) of TRAP-positive multinucleated osteoclasts from FAS-RANK expressers treated with M-CSF and α-FAS (100 ng/ml). The numbers in parentheses show concentrations in nanograms per milliliter. Error bars show averages ± S.D. *, p < 0.05; NS, not significant.