FIGURE 5.

Transfected HDAC4 represses MMP-13 transcription. A, UMR 106-01 cells were transfected with FLAG vector or rat FLAG-tagged HDAC4 construct or no vector. The cells were stimulated with PTH (10⁻⁸ m) for 4 h. Total RNA was extracted from the cells transfected with none (no transfection), FLAG vector, or FLAG-tagged HDAC4. RNAs were measured using Real time RT-PCR. The relative levels of mRNAs were normalized to β-actin and then expressed as fold stimulation over control (cont). The error bars represent ± S.E. of three independent experiments. *, p < 0.05 versus no vector or FLAG vector and treated with PTH for 4 h. B, UMR 106-01 cells were transiently transfected with various vectors (100 ng of pGL2 vector, 100 ng of −148 rat MMP-13 promoter-Luc, 100 ng of FLAG vector, and 100, 200, or 500 ng of FLAG-tagged rat HDAC4), and the luciferase activities were measured with (P) or without (C) 6 h of 10⁻⁸ m PTH treatment. The luciferase activities were normalized to the amount of total protein. The error bars represent ± S.E. of three independent experiments. *, p < 0.05 versus FLAG control. #, p < 0.02; ##, p < 0.01 versus FLAG PTH 6 h. C, UMR 106-01 cells were transfected with FLAG vector or rat FLAG-tagged HDAC4 construct or no vector (none). The nuclear extracts from cells were subjected to immunoblotting with anti-FLAG antibody. Anti-Cdk2 was used as a loading control.