FIG. 2.

(A) HDAC1 and HDAC2 repress TNF-induced expression of a transiently transfected 3XκB-Luc reporter gene. Cos-7 cells were transfected in triplicate with the wild-type 3XκB-Luc reporter and a control vector (pcDNA3.1), pcDNA-HDAC1, pcDNA-HDAC2 alone, or HDAC1 and -2 together. Twenty-four hours after transfection the cells were either untreated or treated with 10 ng of TNF-α/ml for 8 h. Extracts were prepared and relative luciferase activities were determined by normalizing to total protein. Activity is expressed as fold activation relative to the untreated control and is the average ± SD from a representative experiment. Experiments were performed a minimum of three times, with similar results. (B) NIH 3T3 cells harboring either integrated wild-type 3XκB-Luc (left) or mutant 3XκB-Luc (right) were transfected in triplicate as described for panel A. Twenty-four hours after transfection the cells were either untreated or treated with 10 ng of TNF-α/ml for 8 h. Extracts were prepared and relative luciferase activities were determined by normalizing to total protein. Activity is expressed as fold activation relative to the untreated control and is the average ± SD from a representative experiment. Experiments were performed a minimum of three times, with similar results.