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. 2020 Mar 6;11(11):825–845. doi: 10.1007/s13238-020-00701-1

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© The Author(s) 2020

Open AccessThis 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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ERK5 regulates the polymerization of F-actin. (A) Ontological categories of differentially expressed proteins in ERK5-overexpression A549 cells by Metacore^TM GeneGo Pathway Maps analysis. (B) Protein networks associated with the actin regulators differentially expressed in ERK5-overexpression A549 cells (higher definition image is shown in Supplementary file 2: Fig. S4). The network was generated by a shortest paths algorithm of MetaCore^TM (GeneGo) software using the list of differentially expressed actin regulators identified by proteomics analysis. (C) Western blot analysis of actin nucleation and polymerization correlated proteins in ERK5-overexpression A549 cells, including ERK5, Gelsolin, N-WASP, p-PLK1and SPA1. (D) The cell shapes of A549 cells overexpressing ERK5 under light microscope. (E) ERK5 regulated the assembly of F-actin. Immunofluorescence was carried out to display F-actin (phalloidin, red), and nuclei (Hochest, blue) in ERK5-overexpression A549 cells and control A549 cells. (F) After serum starvation overnight, the cells were stimulated again with 20% fetal calf serum for 2 h. Then immunofluorescence assay was performed as that described in (E)