Figure 2

IN is acetylated by GCN5 in vivo. (A) Extracts from HEK 293T cells transfected with the indicated plasmids were immunoprecipitated using anti-Flag antibody and analyzed by Western blotting with anti-acetyl-lysine antibody (upper panel) or anti-Flag antibody (middle panel). Lower panel: cell extracts immunoblotted with anti-HA antibody. (B) Acetylated BSA and peptides corresponding to IN amino acids 260-281, either chemically acetylated at lysines 264, 266, and 273, or not acetylated, were blotted onto a nitrocellulose filter and incubated with anti-acetylated IN antibody. (C) Left panels (lanes 1-4): extracts from HEK 293T cells transfected with the indicated plasmids were immunoprecipitated using anti-Flag antibody and analyzed by Western blotting with anti-acetylated IN antibody (top panel) or anti-Flag antibody (bottom panel). Right panels (lanes 5-8): extracts from HEK 293T cells transfected with the indicated plasmids analyzed by Western blotting with anti-acetylated-IN antibody (top panel) or anti-Flag antibody (bottom panel). (D) Extracts from HEK 293T cells transfected with the indicated plasmids analyzed by Western blotting with anti-acetylated-IN antibody (upper panel), anti-Flag antibody (middle panel), or anti-HA antibody (lower panel).