Fig. 1

Human MAG is a new receptor for ANGPTL2. A Flow cytometric analysis of conditioned medium with/without ANGPTL2-Flag binding to 293T cells with transient transfection of plasmids of N1-hMAG-EGFP (human MAG, full-length) or N1-EGFP (vector). Percentage of binding/unbinding cells to ANGPTL2 in EGFP⁺ cells are shown. B Quantitative data of percentage of binding cells in Panel A (n = 3). C ANGPLT2 bound to the ECD of human MAG (human MAG-ECD-FC) in the conditioned medium (CM) of cotransfected 293T cells using co-immunoprecipitation. Tie-2-ECD-FC served as a negative control. D Representative flow cytometric analysis showing that purified ANGPTL2-Flag protein enhanced GFP expression in human MAG chimeric reporter cells (Human MAG). Control reporter cells (control) without human MAG-ECD were examined. E Quantitative data in Panel D (n = 3). F Binding kinetics of ANGPTL2-Flag to human MAG-ECD were measured using a biolayer interferometry instrument (Octed RED 96/96S). G–H Flow cytometric plots for the binding of ANGPTL2-Flag to full-length, mutant IgG 3 domain or IgG 4 domain of human MAG expressed 293T cells. Percentage of binding/unbinding cells to ANGPTL2 in EGFP⁺ cells (G) and quantitative data are shown (H) (n = 3). (***p < 0.001)