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Fig. 6. Real-time RT-PCR for IL-11 expression in the skeletons of wild-type and Runx2–/– embryos and upregulation of IL-11 expression in Runx2–/– chondrocytes by adenoviral infection of Runx2. (A) IL-11 expression in the skeletons of wild-type embryos (+/+) at E12.5, E13.5, E16.5, and E18.5, and Runx2–/– embryos (–/–) at E18.5. IL-11 expression was examined by real-time RT-PCR. The value in the E12.5 wild-type embryos was defined as 1 and relative values are shown. Data represent the mean of three to five embryos in each group. The IL-11 expression in Runx2–/– skeletons at E18.5 is significantly lower than those in wild-type skeletons at all embryonic stages. P<0.05 as determined by one-way ANOVA. (B) Runx2 and IL-11 expression in Runx2–/– chondrocytes infected with either EGFP-expressing (open columns) or Runx2-and-EGFP-expressing (closed columns) adenovirus. Infected cells were harvested at the indicated times after the onset of viral infection, and Runx2 and IL-11 expression were examined by real-time RT-PCR. The value of Runx2 or IL-11 in Runx2-and-EGFP-expressing adenovirus infection at 48 hours was defined as 1, and relative values are shown. Data represent the mean of three wells. *P<0.05 and **P<0.01 as determined by one-way ANOVA. Similar results were obtained in three independent experiments, and representative data are shown. (C) Synergistic induction of IL-11 mRNA by Runx2 and TGF-ß. Runx2–/– chondrocytes were infected with EGFP-expressing (GFP) or Runx2-and-EGFP-expressing (Runx2) adenovirus. Runx2–/– chondrocytes and Runx2 overexpressing Runx2–/– chondrocytes were treated with TGF-ß (1 ng/ml) for the indicated times. The value of IL-11 in Runx2-and-EGFP-expressing adenovirus infection at 8 hours was defined as 1, and relative values are shown. The induction of IL-11 mRNA by Runx2 was much weaker than that by TGF-ß but Runx2 hugely induced IL-11 mRNA in the presence of TGF-ß. Data represent mean of four wells. *P<0.01 and **P<0.001 as determined by one-way ANOVA. Similar results were obtained in three independent experiments and representative data are shown.
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