Fig. 4: 4E2 activates Tie2 signaling, similar to ANGPT1. | Experimental & Molecular Medicine

Fig. 4: 4E2 activates Tie2 signaling, similar to ANGPT1.

From: An agonistic anti-Tie2 antibody suppresses the normal-to-tumor vascular transition in the glioblastoma invasion zone

Fig. 4

a FACS plots showing the binding capacity of 4E2 to human (left) and mouse Tie2 receptors (right). b Outline of the OCTET competitive binding assay (upper) showing the binding of the 4E2 antibody to immobilized Tie2 followed by competitive binding with ANGPT1 and ANGPT2. Representative binding curves showing the competition modes of ANGPT1 and ANGPT2 with 4E2 (lower). c, d Binding curves showing the competitive binding of 4E2 with ANGPT1 (c) and ANGPT2 (d) to Tie2. e Immunoblots of phosphorylated Tie2 (pTie2) and FoxO1 (pFoxO1) in HUVECs treated with 4E2 and quantitative analysis (n = 5 per group). *, P < 0.05; **, P < 0.01 vs. 0 μg/mL 4E2. f Immunofluorescence images of FoxO1 in HUVECs and the proportions of its cytosolic and nuclear localization. The arrows indicate nuclear exclusion of FoxO1 (n = 3 per group). *, P < 0.05; **, P < 0.01 vs. Ctrl of nuclear FoxO1. #, P < 0.05; ##, P < 0.01 vs. Ctrl of cytosolic FoxO1. g Immunofluorescence images and quantification of junctional VE-cadherin in HUVECs (n = 5 per group). h Quantification of permeability in HUVECs. Scale bars: 100 µm (f), 25 µm (g). ns not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001.

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