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. 2021 May 12;40(23):3929–3941. doi: 10.1038/s41388-021-01798-2

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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 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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Fig. 3 — A, B Immunoblot analysis confirmed that shERK5(UTR) and shERK5(CDS) effectively downregulated ERK5 expression in MDA-MB-231 cells. Ectopic expression of ERK5 [+ F-ERK5(WT)] at a level similar to that of the endogenous protein was achieved by incubating shERK5(UTR)-expressing cells carrying an inducible F-ERK5(WT) construct with 10 ng/ml doxycycline (+) for 24 h. C ERK5 knockdown suppressed MDA-MB-231 cell invasion in FBS-free medium through Matrigel. The bar graph indicates the mean ± SD (N = 3). Representative pictures of the filters are shown. Scale bar: 200 μm. D Quantitative time course analysis of MDA-MB-231 cell adhesion on Matrigel by IncuCyte® live-cell imaging system. Representative pictures of cells taken 4 h after seeding in Matrigel-coated plates are shown. The data correspond to the mean ± SD (N = 3). E Lysates were obtained from cells cultured in FBS-free Matrigel-coated plates for 12 h and analyzed by immunoblot. The images are representative of three biological repeats. F Alternatively, transcript levels were measured by quantitative real-time PCR. Results are presented as fold change ± SD. The graph is representative of three independent biological repeats performed in duplicate.