JMJD6 is physically associated with p53 <i>in vivo</i> and <i>in vitro</i>.

<p>(A) Cellular extracts from HCT116 cells stably expressing vector or FLAG-JMJD6 were immunopurified with anti-FLAG affinity columns and eluted with FLAG peptide. The eluates were resolved by SDS-PAGE and silver-stained. The proteins bands were retrieved and analyzed by mass spectrometry. (B) HCT116 cell lysates were immunoprecipitated with antibodies against JMJD6 followed by immunoblotting with antibodies against p53 (FL-393), or they were immunoprecipitated with antibodies against p53 (FL-393) followed by immunoblotting with antibodies against JMJD6. (C) Immunofluorescence-stained endogenous JMJD6 (red) and p53 (green) were visualized by confocal microscopy. DAPI staining was included to visualize the cell nucleus (blue). Scale bar, 25 µm. (D) Mapping the domain of p53 that is required for its interaction with JMJD6. GST pull-down experiments were performed with GST-fused JMJD6 and <i>in vitro</i> transcribed/translated Myc-tagged full-length p53 or deletions of p53.