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. 2020 Jan 3;19(3):1789–1796. doi: 10.3892/etm.2020.8419

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Copyright: © Chen et al.

This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

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Figure 2. — Inhibition of EphA4-VEGFR2 binding by EphA4(ΔJM, KD)-GFP. (A) EphA4-Flag and VEGFR2-HA were co-expressed with increasing doses of EphA4(ΔJM, KD)-GFP in 293T cells, and the binding of EphA4-Flag and VEGFR2-HA was examined using IB with or without IP using the antibodies shown following SDS-PAGE. (B) Inhibition of ligand-mediated receptor phosphorylation by EphA4(ΔJM, KD) tagged with GFP. EphA4-Flag and VEGFR2-HA were co-expressed in 293T cells with or without EphA4(ΔJM, KD)-GFP, the phosphorylation of VEGFR2 was examined using IP with anti-VEGFR2 followed by IB with anti-pY following 0 or 10 min stimulation with 20 ng/ml VEGF165. ΔJM, juxtamembrane domain deleted; Eph, ephrin receptor; GFP, green fluorescent protein; HA, hemagglutinin; IB, immunoblotting; IP, immunoprecipitation; KD, kinase-dead; pY, phosphotyrosine; VEGF, vascular endothelial growth factor; VEGFR, VEGF receptor; WT, wild-type.