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Fig. 1. Hint1 directly binds to Pontin and Reptin. Purified recombinant GST-Pontin, -Reptin, -ß-catenin, -LEF-1 and -Hint1 fusion proteins were incubated with MPB-Hint1. Protein complexes were pulled down with GSH-agarose beads and analysed by western blotting with anti-MBP and anti-GST antibodies. MBP-Hint1 specifically binds to GST-Pontin (lane 4) and GST-Reptin (lane 6) but not to GST (lane 2), GST-ß-catenin (lane 8) or GST-LEF-1 (lane 10). GST-Hint1 associates with MBP-Hint1 (lane 12) as expected from the crystal structure published previously. In control experiments GST and GST-fusion proteins do not nonspecifically bind to MBP or MBP-Hint1 (lanes 1,2,3,5,7,9,11).





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