FIG. 2.

JMJD2A is a nuclear protein that possesses transcriptional-repression activity. (A) Immunostaining revealed that JMJD2A is primarily a nuclear protein in HeLa cells. Also shown is DAPI (4′,6′-diamidino-2-phenylindole) staining of nuclei. (B) JMJD2A functioned as a repressor when tethered to a target reporter. The full-length JMJD2A was fused to the Gal4 DNA binding domain (Gal-JMJD2A), and repression was measured by cotransfection with the 4xUAS-TK-luc reporter. The expression of Gal-JMJD2A was confirmed by Western analysis, as shown at the bottom of panel. (C) Repression by Gal-JMJD2A was correlated with recruitment of N-CoR and SMRT corepressor complexes. HeLa cells were transfected with reporter alone or reporter plus Gal(DBD) or Gal-JMJD2A, and, after overnight incubation, ChIP assays were carried out to determine the recruitment of SMRT and N-CoR and their associated HDAC3 to the reporter.