The interacting regions of nectin-4 and the prolactin receptor.
A, the region of nectin-4 interacting with the PRLR. Aa, schematics of the domain structures of nectin-4 and its mutants. Ab, identification of the PRLR-interacting region of nectin-4. HEK293E cells were transfected with various combinations of the indicated plasmids and cultured in suspension. The FLAG-tagged nectin-4 or each of its mutants was immunoprecipitated from each lysate using the anti-FLAG Ab. Each sample was subjected to Western blotting using the indicated Abs. B, the region of the PRLR interacting with nectin-4. Ba, schematics of the domain structures of the PRLR and its mutants. Bb, identification of the nectin-4-interacting region of the PRLR. HEK293E cells were transfected with various combinations of the indicated plasmids and cultured in suspension. The HA-tagged PRLR or each of its mutants was immunoprecipitated from each lysate using the anti-HA Ab. Each sample was subjected to Western blotting using the indicated Abs. The results are representative of three independent experiments. SP, signal peptide; TM, transmembrane; CP, cytoplasmic; IB, immunoblotting; IP, immunoprecipitation.