In vivo and in vitro interaction of GAS41 and INI1.

(A) Immunoblot analysis with a goat polyclonal anti-INI1 antibody of GAS41 protein immunoprecipitated from whole extract of KG1a cells (lane 1); 30 μg protein extract was used as antibody positive control (lane 2). Immunoprecipitates with rabbit preimmune serum were used as negative control (lane 3). Samples were run on a 12% polyacrylamide gel. (Bi) Autoradiograph of a 12% polyacrylamide gel with in vitro–translated INI1 protein precipitated with 2 different GST-GAS41 fusion polypeptides. Lane 1, input of in vitro–translated INI1 protein (one fifth of total amount); lane 2, GST-GAS41–AF9-like region (aa 1-95); lane 3, GST-GAS41–coil-coiled region (aa 163-227); and lane 4, GST as negative control. (Bii) The equivalent Coomassie blue–stained gel of the pull down experiment. (C) Immunoblot analysis with a goat polyclonal anti-INI1 antibody of AF10 protein immunoprecipitated from whole extract of KG1a cells (lane 1). Protein extract (30 μg) was used as antibody positive control (lane 2). Immunoprecipitates with rabbit preimmune serum were used as negative control (lane 3). Samples were run on a 12% polyacrylamide gel.