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The protein expression of BSP and OC in cellular lysates was analyzed by Western blot analysis. Dex significantly increased the protein level of BSP from day 1 to day 6 when compared to Dex-untreated MLN9708 (Fig. 2B). OC protein was also increased in Dex-treated cells compared to Dex-untreated cells at day 6.
Effect of Dex on Runx2 expression and transcriptional activity
Fig. 3.
Effect of Dex on the expressions and transcriptional activity of Runx2 in ROS17/2.8 cells. (A) mRNA expression of Runx2. Cells were cultured with or without treatment of 10?8 M Dex for 24 h, or pretreated with 10 μg/ml cycloheximide for 30 min, and then 10?8 M Dex was added for an additional 24 h. mRNA expressions were determined by real time RT-PCR. (B) Protein expression of Runx2. Cells were cultured with or without treatment of 10?8 M Dex for 1, 3, and 6 days. The immunoblot was probed with the anti-Runx2 antibody. (C) Transcriptional activity of Runx2. A promoter construct, (OSE2)x6-Luc, was transfected into cells. After 24 h, cells were treated with vehicle (ethanol), 10?8 M Dex, 10?6 M RU486, or 10?8 M Dex + 10?6 M RU486 for an additional 24 h, and then luciferase activity was determined. A vector pGL-Basic (Basic) was used as negative control.
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