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. Author manuscript; available in PMC: 2023 Jul 21.
Published in final edited form as: Cell Chem Biol. 2022 Jun 15;29(7):1162–1173.e5. doi: 10.1016/j.chembiol.2022.05.008

Figure 2. NCoR binds to HDAC4, 5, and 7 in an acetyllysine-dependent manner.

Figure 2.

Flag-tagged HDAC4 (A), HDAC5 (B), and HDAC7 (C) were overexpressed in HEK293 cells, which were then treated with SAHA (10 μM) to induce acetylation. After lysis, anti-Flag beads were used to immunoprecipitate (IP) the HDAC proteins either in the absence or presence of 1 mM LGK (K), LGKAc (KAc), or SAHA (SA). Bound proteins from the IP were resolved by SDS-PAGE, followed by Western blot analysis with NCoR and Flag antibodies. Lysate (Lys) from transfected cells were included as a gel migration control. All trials include a bead binding control using lysates without expression of HDAC-Flag (IgG). Additional independent trials and dose dependence are shown in Figure S3A–C. Binding affinity studies are shown in Figure S4. (D) NCoR band intensities from three independent trials with HDAC4, 5 and 7 (parts A-C and S73A-C) were quantified and normalized to untreated (set to 100%), with the student t test applied to show significance. N.S. = not significant (p > 0.05), and ** = p < 0.01. Raw data are shown in Figure S3D.