Extended Data Fig. 3: EphB4 interacted with InsR.
From: Insulin induces insulin receptor degradation in the liver through EphB4
a, The identified peptide from EPHB4 by mass spectrum in INSR interaction protein screening in HepG2 cells. The b- and y-type product ions are marked on the mass spectrum and also illustrated in the peptide sequence shown above. b, c, Expression pattern of EphA and EphB families in C57BL/6 J mice by semiquantitative PCR. d, Interactions of homologous proteins of EPHB4 with InsR in HepG2 cells. e, Interaction of EPHB4 with other tyrosine kinases in HepG2 cells. f, Interaction of EPHB4 with IGF1R in HepG2 cells. g, h, Interaction of EPHB4-HA and Flag-INSR in HEK293T cells after Flag-INSR or EPHB4-HA was immunoprecipitated. i, Co-localization of EPHB4 and INSR detected using endogenous antibodies. Pearson’s R value: 0.57. j, Direct binding of EPHB4 and InsR in a pull-down assay using purified proteins. k, Diagrammatic sketch of the InsR protein structure. The short line indicates the truncated mutants of the full-length InsR protein. l, Interaction of EPHB4 with INSR domains with INSR truncation mutants. m, Diagrammatic sketch of the EPHB4 protein structure. The short line indicates the mutants in which part of the sequence was deleted from the full-length EPHB4 protein. n, Interaction of INSR with EPHB4 domains in EPHB4 deletion mutants. o, p, Interaction of INSR with EPHB4 (WT) and its kinase-dead mutant (KD) in HepG2 cells. n = 3 independent experiments in p. For statistical analysis in p, comparisons were versus related controls, using two-sided Student’s t-test. Values are presented as the mean ± SEM.
