SIRT1 interacts with and deacetylates FOXA2.

<p>(A) Increase in FOXA2 acetylation by NAM. FLAG-FOXA2-transfected HEK293T cells were treated with class I/II HDAC inhibitor TSA (5 µM), class III HDAC inhibitor NAM (20 mM) or both for 16 hours. Acetylation level of immunoprecipitated FLAG-FOXA2 was assessed on Western Blot probed with an anti-acetylated-lysine specific antibody. (B) SIRT1 co-transfection affects FOXA2 acetylation. HEK293T cells were transfected with FLAG-FOXA2 alone or together with MYC-SIRT1 in the presence of 20 mM NAM. Acetylation was assessed as in (A). (C) SIRT1 interacts with FOXA2. HEK293T cells were co-transfected as in (B) followed by co-immunoprecipitation with anti-FLAG M2 beads. Interaction of FLAG-FOXA2 and MYC-SIRT1 was assessed by Western Blot analysis with indicated antibodies. (D) Endogenous FOXA2 and SIRT1 interact in the nucleus. HepG2 cells were subjected to immunofluorescence-based proximity ligation assay <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0098438#pone.0098438-Plas1" target="_blank">[28]</a>. Nuclei were counterstained with DAPI.