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. 2020 Nov 4;11:5574. doi: 10.1038/s41467-020-19211-z

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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. 7 — a HCT-116 cell viability upon treatment with increasing concentrations of bis-ANS for 6 or 24 h as determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Error bars represent SD (n = 5 technical replicates across two biological replicates). b Live-cell imaging of HCT-116 cells with or without 1 h treatment with 250 μΜ bis-ANS. Nuclei were stained with 1:1000 Draq5. c Schematic diagram of sodium arsenite washout experiments described. HCT-116 cells were treated for 30 min with 500 μΜ sodium arsenite in culture medium. Arsenite was removed and medium containing 250 μΜ bis-ANS or medium alone (control) was added. Cells at different time points after sodium arsenite removal were fixed and stained with DAPI and with an antibody against G3BP. d Representative microscopy images obtained 60 and 90 min after arsenite removal. e Quantitative assessment of percentage of cells exhibiting punctate G3BP staining in the presence and absence of bis-ANS at different times after arsenite removal. Note that data points at 0 min represent analysis of the same images for control and treatment groups. Scale bars represent 20 μm. All data are presented as mean values ± standard deviation (n = 3, each independent experiment analyzing 20 fields of view).