Figure 7.

Increased nuclear localization of HDAC4 enhances MEF2-dependent transcriptional repression. TAg-Jurkat cells were transfected with MEF2D, a MEF2-luciferase reporter construct, a constitutive β-gal expression construct, and either wild-type HDAC4 or HDAC4 with mutations in all three 14-3-3 binding sites (HDAC4 S246/466/632A). pcDNA3.1 (Invitrogen) was used to normalize the amount of DNA transfected. Thirty-eight hours after transfection, the samples were harvested and divided to perform the subsequent assays in triplicate. The amount of luciferase activity was measured (Promega) and divided by the amount of β-gal activity present to normalize for protein expression levels.