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. 2020 Jan 3;11:74. doi: 10.1038/s41467-019-13771-5

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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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Fig. 2 — a A schematic explanation of the experiment to identify the acute response of ALK-rearranged cells to alectinib. Four ALK-rearranged NSCLC cells with alectinib sensitivity were exposed to DMSO or the survivable inhibitory concentration (sIC) of ALC (see Table 1) for 96 h. Proteome and conventional biological analyses were performed to compare cells that survived ALC and proliferating control cells (DMSO-treated). b The integrated results of the proteome analysis in ALK-rearranged cell lines (n = 3 biologically independent cell lines, KTOR3 was excluded, please see the text). Volcano plot representing all expressed proteins. Regarding every protein, fold changes in the vehicle and exposure to alectinib were plotted against the –log P-value. The integrated P-value was calculated by paired t-test. Significant differentially expressed proteins, with a fold change ≥1.5 or ≤−1.5, are depicted as red or blue, while insignificant ones are shown as black dots. Of the 3183 proteins detected after the 96-h exposure to alectinib, 48 were differentially expressed (18 up- and 30 downregulated). c Gene ontology (GO) analysis of the 48 proteins. P-values were calculated using Fisher’s test. d KEGG pathway analysis on the upregulated proteins. P-values were calculated using Fisher’s test. All annotated pathways are presented. e Average cell area on the culture plate. A plot shows the average cell area on each photographed image. Significance was evaluated using a one-way ANOVA followed by Sidak’s multiple comparison test. f Cell adhesion areas in culture plates increased when ALK-rearranged cells were treated with ALC. g A simplified diagram of the Hippo signaling pathway using KEGG pathway analysis. Two important factors (Ajuba, AFP) among the four annotated proteins are shown. All analysis results are described in Supplementary Data 2. *P < 0.05. ALK anaplastic lymphoma kinase, NSCLC non-small cell lung cancer, ALC alectinib, DMSO dimethyl sulfoxide. Error bars indicate ± S.D.