FIG. 6.

Repression ability of HDAC4 and its mutant S246/467/632A. (A and B) The reporter (200 ng), MEF2-E4-Luc (A) or pJLuc (B), was transfected into NIH 3T3 cells with a MEF2C expression plasmid (100 ng), an internal control plasmid (CMV-β-Gal; 50 ng), and the expression plasmid for Flag-tagged HDAC4 or S246/467/632A at the indicated amount. The normalized luciferase activity from the transfection without any effector plasmid was arbitrarily set to 1.0. Average values of at least three independent experiments are shown with standard deviation indicated by error bars. (C) A 200-ng sample of the Gal4-tk-Luc reporter was transfected into NIH 3T3 cells with a Gal4-VP16 expression plasmid (5 ng), the internal control plasmid CMV-β-Gal (50 ng), and the expression plasmid for Flag-tagged HDAC4 or S246/467/632A at the indicated amount. The reporter activities were measured as for panels A and B. (D) The Gal4-tk-Luc reporter was transfected into NIH 3T3 cells along with an expression plasmid for Gal4-HDAC4 or Gal4-S246/467/632A. Normalized luciferase activities from transfection with effector plasmids at the indicated amounts were compared with that from the reporter alone to calculate the relative repression. Average values of four independent experiments are shown with standard deviation indicated by error bars.