Fig. 3.

AP4–Gem–SMRT complex interacts with HDAC3 to establish and maintain TSA-dependent repression of its target gene in nonneuronal cells. (A) AP4–Gem–SMRT complex was immunoprecipitated with anti-AP4 antibody in HEK293T cells, eluted away from the antibody, and then immunoprecipitated again with an anti-SMRT antibody. The final precipitate was probed for Gem. (B) HDAC3 IP was performed in HEK293T cells, and Gem, AP4, and SMRT subsequently were analyzed by immunoblotting. (C) Increasing doses of Gem were transiently cotransfected with synthetic GAL4-Gem construct in vehicle- or TSA (200 nM)-pretreated HEK293T cells, and increasing doses of HDAC3 were transiently cotransfected with or without 30 ng of Gem along with synthetic GAL4-Gem construct in HEK293T cells. Values are mean ± SEM (n = 3). ∗, P < 0.05. (D) Time course for derepression of the PAHX-AP1 and DYRK1A genes in HEK293T cells after exponentially proliferating cells were incubated in the presence of TSA.