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. 2009 Oct 12;423(Pt 3):389–400. doi: 10.1042/BJ20090086

Figure 1. HDAC2 is critical for binding of APPL1 to the NuRD complex.

Figure 1

(A and B) Extracts from HeLa cells transfected for 72 h with two (a, b) or three (a, b, c) different siRNA oligonucleotides per gene against: HDAC1, HDAC2, MTA2, RbAp48 and RbAp46 or non-specific siRNA (Φ) were subjected to immunoprecipitation (IP) using: (A) anti-APPL1 antibody; (B) anti-HDAC1 antibody (left panel) or anti-MTA2 antibody (right panel). Non-specific antibodies (IgG) were used as controls. Input indicates 10% of total cell extracts used for immunoprecipitation. Immunoprecipitates and input extracts were analysed by Western blotting using different antibodies as indicated. (C) To verify the direct interactions between APPL1 and HDAC1 or HDAC2, in vitro translated HDAC1–FLAG and untagged HDAC2 were subjected to GST pull-down assay using GST alone (Φ) or GST fused to the N- or C-terminal parts of APPL1 (APPL1-N or APPL1-C, respectively). Input indicates 10% of in vitro translated material used for the pull-down assay. Bound proteins were analysed by Western blotting using anti-HDAC1 and anti-HDAC2 antibodies. ND, not determined.