Fig. 5. Osteoclastogenesis is affected in cells from JunAA/JunAA mice and in cells lacking c-Jun but not lacking JunD. (A) Co-culture of JunAA/JunAA or wild-type hematopoietic precursors with wild-type osteoblasts. (B) Osteoclast differentiation of bone marrow precursors treated with M-CSF (20 ng/ml) and RANKL (50 ng/ml), or M-CSF (20 ng/ml) and TNF- (50 ng/ml) (C). The data represent results obtained from two experiments (B) or with pairs of littermate mice (C). (D) The function of JunD in osteoclastogenesis was addressed by direct stimulation of bone-marrow precursors isolated from wild-type, JunD^+/- or JunD^-/- mice by M-CSF and RANKL. (E) RANKL-mediated differentiation of double mutated cells JunD^-/-; JunAA/JunAA compared with JunD^+/-; JunAA/JunAA cells. (F) RANKL-mediated differentiation of BMMs lacking c-Jun (/) compared with wild-type (wt) or heterozygous controls (/wt). Osteoclast differentiation was monitored by TRAP staining from day 3 to 6 (A) or at day 4 (C,D,E,F) of culture. The deletion of c-Jun was measured by PCR analysis of DNA isolated from the cultured cells (data not shown). The stars indicate the statistical significance: (^***) P<0.0001, (^**) P<0.001; (ns) non-significant.

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