In vitro interaction of AF10 and GAS41.

(Ai) Autoradiograph of a 12% polyacrylamide gel with in vitro–translated GAS41 protein pulled down with 4 different GST-AF10 fusion polypeptides. Lane 1, input of in vitro–translated GAS41 protein (one fifth of total amount); lane 2, GST-AF10–cysteine-rich region (aa 1-331); lane 3, GST-AF10–leucine zipper motif (aa 733-806); GST-AF10–leucine zipper region (aa 683-972); lane 5, GST-AF10–C-terminal portion (aa 819-972); and lane 6, GST as negative control. (Bi) Autoradiograph of a 10% polyacrylamide gel with in vitro–translated AF10 protein precipitated with 2 different GST-GAS41 fusion polypeptides. Lane 1, input of in vitro–translated AF10 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. (Ci) Autoradiograph of a 12% polyacrylamide gel with in vitro–translated GAS41 (lane 1, input of in vitro–translated protein), precipitated with the leucine zipper motif of AF17 (aa 696-769), lane 2; and with GST in lane 3 as negative control. (Aii,Bii,Cii) The right panels represent the equivalent Coomassie blue–stained gel of the pull-down experiment.