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. 2012 Feb 13;209(2):319–334. doi: 10.1084/jem.20111566

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© 2012 Zhao et al.

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Figure 8. — RBP-J maintains expression of IRF-8, which suppresses osteoclastogenesis. (A) Immunoblot analysis of IRF-8 expression in Rbpj^+/+ and Rbpj^ΔM/ΔM BMMs obtained at the indicated time points after stimulation with TNF. p38 was measured as a loading control. (B) Immunoblot analysis of IRF-8 in control and NICD^M BMMs stimulated with RANKL for the indicated times. (C) Immunoblot analysis of IRF-8 in control and NICD1^M BMMs transfected with Rbpj or control siRNAs. p38 was measured as a loading control. (D) Immunoblot analysis of IRF-8 in Rbpj^ΔM/ΔM BMMs transduced with the retroviral vectors pMX-IRES-EGFP (Control) or pMX-Irf8-IRES-EGFP (Irf8). p38 was measured as a loading control. (E) Osteoclastogenesis assays using Rbpj^ΔM/ΔM BMMs transduced with the retroviral vectors pMX-IRES-EGFP or pMX-Irf8-IRES-EGFP without or with TNF stimulation. The number of GFP-positive MNCs (osteoclasts) per well was counted. **, P < 0.01. (F) Fluorescent images of the Rbpj^ΔM/ΔM BMMs transduced with the retroviral vectors pMX-IRES-EGFP or pMX-Irf8-IRES-EGFP with TNF stimulation. The GFP-positive multinucleate cells (osteoclasts) appear as large green cells. Bar, 100 µm. Data are representative of at least three independent experiments (A–F).