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. 2009 Apr 22;284(27):18174–18183. doi: 10.1074/jbc.M109.001354

FIGURE 9.

FIGURE 9.

MN1 stimulates RANKL promoter activity. A, MC3T3-E1 cells were transiently transfected with a 1-kb human RANKL promoter reporter gene (hRANKL(−1kb)-luc) or pGL3 control reporter gene. The indicated amounts of the MN1 expression plasmid were cotransfected with the total molar amount of expression plasmid DNA kept constant by the addition of empty vector. The normalized luciferase activity was measured after 48 h. Data represent the mean ± S.D. (n = 3 in each group). B, control siRNA (−) or MN1 siRNA (+) was transfected into MC3T3-E1 cells 24 h before pGL3 or hRANKL(−1kb)-luc was transfected. The normalized luciferase activity was measured 48 h later. Data represent the mean ± S.D. (n = 3 in each group). The inset shows a Northern blot analysis of MN1 mRNA expression 24 h after siRNA transfection. C, WT or MN1 KO primary osteoblasts were transfected with pGL3 or hRANKL(−1kb)-luc. The normalized luciferase activity was measured after 48 h. Data represent the mean ± S.D. (n = 3 in each group). D, WT or KO primary osteoblasts were transiently transfected with pGL3 or hRANKL(−1kb)-luc. The indicated amounts of the MN1 expression plasmid were cotransfected with the total molar amount of expression plasmid DNA kept constant by the addition of empty vector. The normalized luciferase activity was measured after 48 h. Data represent the mean ± S.D. (n = 3 in each group).