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. 2021 Apr 23;11:8856. doi: 10.1038/s41598-021-88382-6

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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EphA2 activation by ephrinA5 results in weaker canonical EphA2-pY588 signaling than upon ephrinA1-mediated activation. (A–C) EphA2 (total and phosphorylated at Y588 or S897) in OVCAR3 and OVCAR4 after treatment with soluble recombinant dimeric (Fc-tagged) and monomeric (His-tagged) ephrinA1 and ephrinA5 for 120 min were assessed by immunoblotting (A) and quantified for EphA2-pY588 (B) and EphA2-pS897 (C). N = 3. Full-length blots are presented in Supplementary Fig. 5. (D, E) Fluorescent micrographs of EphA2 (red) and EpCAM (green) in ephrinA1-Fc and ephrinA5-Fc treated OVCAR3, OVCAR4, and OVCAR8 cells (D) and corresponding EphA2 quantification (E). N = 3 (OVCAR3) or 4 (OVCAR4 and OVCAR8). Mock is set to one. p values (Student’s t-test): ns = not significant, *< 0.05; **< 0.01; ***< 0.001.