Fig. 3. MCEMP1-KIT interaction amplifies downstream signal transduction.

a Co-immunoprecipitation assay of human and mouse MCEMP1 and KIT interaction in 293 T cells. b Co-immunoprecipitation assay of MCEMP1 interaction with wild-type KIT or enzymatic dead (D794N) mutant in 293 T cells. c Schematic diagram of MCEMP1 and KIT structure and co-immunoprecipitation assay of MCEMP1 interaction with KIT deletion mutants in 293T cells. ΔKI was not expressed. d Immunoprecipitation and Immunoblot analysis of MCEMP1 and KIT phosphorylation in C57 cells expressing vector (VEC), wild-type MCEMP1 (WT), or YF mutant MCEMP1 (YF). C57 cell lysates were immunoprecipitated with anti-Flag antibody. Immunoprecipitates and WCL were analyzed by IB with the indicated antibodies. Band intensity of Immunoprecipitated KIT and phosphorylated KIT was measured with densitometric analysis by ImageJ and normalized to the intensity of MCEMP1 WT or YF. Data are presented by min to max of box and whiskers and p-values were determined by two-tailed unpaired Student’s t-test (n = 4). e KIT phosphorylation and downstream MAPK signal transduction upon SCF stimulation in C57 cells expressing VEC, WT MCEMP1 or YF mutant MCEMP1. f, g Gene expression of Il6 and Il13 after SCF stimulation in C57 cells expressing VEC, WT MCEMP1 or YF mutant MCEMP1. Data are representative of at least two independent experiments in a–e. Data are presented by mean±s.e.m. and p-values were determined by two-way ANOVA with Sidak’s multiple comparison in f (n = 3) and g (n = 3).