Fig. 1

Characterization of EGFR/TAM chimeric receptors. a Schematic representation of wild-type TAM receptors. The ligands, (Gas6 and Pros1) serve as bridging molecules to link TAMs on phagocytic cell to externalized PS on apoptotic cells. b Schematic representation of EGFR/TAM chimeric receptors created by fusing the extracellular domains human EGFR with the trans-membrane and intracellular domains of each TAM receptor. c The percentage identity between the different domains of TAMs and the different tyrosine-based motifs on TAMs intracellular kinase domain that can be phosphorylated (the asterisks indicate the autophosphorylation sites). d EGFR expression on stable EGFR/TAM cell lines as analyzed by flow cytometry. e-g Immunoblots analysis of stable EGFR/Tyro3 (e), EGFR/Axl (f) and EGFR/Mertk (g) CHO cell lines characterizing receptor expression, and EGF-inducible dimerization and activation of functional proteins verified using pTyro3 (e), pAxl (f), and pMertk (g) antibodies. h Representative bright-field micrographs of parental and EGFR/TAM CHO cells seeded on fibronectin-coated (upper panels) or uncoated (lower panels) plastic surface. The Inset shows a representative enlarged single cell. i The cell axial ratio (cell length/width) quantification data of parental and EGFR/TAM CHO cells seeded on fibronectin-coated or plastic surface. Differences between groups were tested by two-way ANOVA and Tukey’s multiple comparisons test, *P < 0.05, **P < 0.01, ****P < 0.0001. j Immunoblot analysis of N-Cadherin induction by TAMs. k Immunoblot analysis showing effects of 200nM R428 and 200nM BMS777607 on TAMs activation. l Densitometry analysis of the immunoblots showing the percentages of inhibition compared to EGF treatment only. Mean values ± SD are shown (n = 3)