LGP2 interaction requires the C-terminal and the transmembrane regions of IPS-1. (A) Schematic representation of IPS-1 fragments used for mapping. Positions of FLAG or GST tags (T), CARD, the proline-rich region (Pro-rich), and the transmembrane domain (TM) are indicated. (B) HEK 293T cells were transfected with expression vectors for six-His-tagged LGP2 and the FLAG-tagged regions of IPS-1, as indicated. Cell lysates were subjected to FLAG immunoprecipitation using M2-agarose beads. His-LGP2 protein copurification with FLAG-tagged IPS-1 was analyzed by immunoblotting, using six-His antibody. (C) HEK 293T cells were transfected with expression vectors for His-tagged LGP2 and GST-tagged regions of IPS-1, as indicated. Cell lysates were subjected to glutathione bead affinity purification. Six-His-LGP2 protein copurification with GST-IPS-1 was analyzed by immunoblotting, using six-His antibody.