Fig 1: Transcriptional repressive activities of Gal4N-fused SRID mutants of HDAC4c.(A) Mammalian one-hybrid assay of HDAC4c mutants. HEK293 cells were cotransfected with the Gal4-tk-luciferase reporter plasmid (200 ng) and wild-type (WT) or the indicated mutants of Gal4N-HDAC4c plasmids (50 and 100 ng), together with the pCMV-β-gal vector (100 ng). After 48 hours of transfection, the luciferase activities were measured and normalized, as described in the materials and methods section. Fold repression indicates the mean ± S. E. value obtained from at least two independent experiments performed in duplicate. The p-values for all compared groups between wild-type and mutants are less than 0.05. RLU: relative luciferase activity. (B) The expression levels of wild-type (WT) or indicated mutants of Gal4N-HDAC4c proteins were examined by immunoblot analysis of whole cell lysates (15 μg) using anti-Gal4N mouse monoclonal antibody (Santa Cruz, sc-510; 1:3,000 dilution).