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. 2020 May 22;10:8537. doi: 10.1038/s41598-020-65250-3

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© The Author(s) 2020

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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ERK isoforms have opposite effects on lung metastatic burden. (A) copGFP-labeled SUM149 SCR, shERK1, and shERK2 cells were generated, and ERK isoform knockdown was confirmed by immunoblotting. (B,C) Nine weeks after cancer cell injection, mice were sacrificed, lung metastatic burden was observed using stereomicroscopy, and total tumor area was quantified. ERK1 knockdown dramatically increased metastatic burden. ERK2 knockdown almost entirely prevented lung metastasis. (D) Micrographs (20X) of lung SUM149 SCR and SUM149 shERK1 tumors immunohistochemically stained for hematoxylin and eosin, proliferation marker Ki67, ERK, and phosphorylated ERK. (E) Kaplan-Meier survival analysis shows that mice bearing SUM149 shERK2 tumors survived longer than control mice or mice bearing SUM149 shERK1 tumors. No tumor tissue was identified in the lungs of mice injected with shERK2 40-44 cells.