Figure 1. HDAC inhibitor-dependent HDAC-NCoR binding.
(A) Flag-tagged wild type (WT) or GOF HDAC4 (A), HDAC5 (B), HDAC7 (C), or HDAC9 (D) were overexpressed in HEK293 cells, which were then treated with SAHA (10 μM) to induce acetylation. After lysis, Flag-tagged HDAC proteins were immunoprecipitated (IP) in the presence or absence of SAHA. Bound proteins were resolved by SDS-PAGE, followed by western blot analysis with NCoR and Flag antibodies. As a gel migration control, lysate (Lys) from transfected cells were included. All trials include a bead binding control using lysates without expression of HDAC-Flag (IgG). Activity assays associated with the wild type and mutant HDAC proteins are shown in Figure S1. Additional independent trials and SAHA dose dependence are shown in Figure S2.