Critical roles of c-Jun signaling in regulation of NFAT family and RANKL-regulated osteoclast differentiation
J. Clin. Invest. Fumiyo Ikeda, et al. 114:475 doi:10.1172/JCI19657 [Go to this article.]

Figure 1
Osteopetrotic phenotype of TRAP-DN–c-Jun TG mice. (A) Expression of DN–c-Jun transgene in TRAP-DN–c-Jun TG mice. Total RNA isolated from brain (lanes 1 and 7), kidney (lanes 2 and 8), liver (lanes 3 and 9), lung (lanes 4 and 10), spleen cells (lanes 5 and 11), or BMMΦ (lanes 6 and 12) of wild-type (WT; lanes 1–6) and TRAP-DN–c-Jun TG (TG; lanes 7–12) littermate mice were subjected to RT-PCR analysis using specific primers for DN–c-Jun (top panel) or β-actin (bottom panel). (B) Appearance of wild-type and TRAP-DN–c-Jun TG littermate mice. No tooth had erupted in TRAP-DN–c-Jun TG mice. (C) Radiological analyses in TRAP-DN–c-Jun TG mice. TRAP-DN–c-Jun TG mice showed increased radiodensity of long bones, vertebrae, and jaw compared with wild-type littermates. (D) H&E and TRAP staining of long bones of WT or TRAP-DN–c-Jun TG littermate mice. Left panels, lower magnification (H&E, ×50; TRAP, ×100); right panels, higher magnification (H&E, ×100; TRAP, ×400). (E) Impaired osteoclastogenesis of TRAP-DN–c-Jun TG mice in vitro. Spleen cells isolated from wild-type or TRAP-DN–c-Jun TG littermate mice were incubated with M-CSF and sRANKL for 6 days, and TRAP⁺ multinucleated osteoclast-like (ocl) cells were counted. (F) Macrophage-like cell differentiation determined by NSE staining in the culture of spleen cells in α-MEM–10% FCS with M-CSF. (G) Osteoblast differentiation determined by ALP staining (left panel) or evaluated for ALP activity (right panel) in the culture of primary osteoblasts isolated from calvariae in α-MEM–10% FCS with BMP2.