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. 2020 Oct 27;3:620. doi: 10.1038/s42003-020-01339-9

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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. 1 — a, b Matrigel chemoinvasion assay of MDA-MB-231 cells (a) and Hs578T cells (b) with or without treatment of lysosomal inhibitors bafilomycin A1 (Baf A1) or chloroquine (CQ) and 4 Gy IR. Representative images of the results were obtained from the Matrigel invasion assays. c Lysosomal exocytosis was quantified by the measurement of dextran-488 in culture medium. d Cell-surface LAMP1 was measured by flow cytometry. e Cell-surface LAMP1 fluorescence intensity was measured using an image scanner. The fluorescence intensity was normalized to the surviving cell viability in each group. f Immunofluorescence images of LAMP1 in MDA-MB-231 cells treated with or without IR treatment. Green, LAMP1; red, α-tubulin; yellow, γ-tubulin (centrosome); blue, DAPI (nucleus). Bar, 10 μm. g Total LAMP1 in whole-cell lysates of MDA-MB-231 cells after IR treatment was detected by immunoblotting. h The lysosomal distribution from the centrosome to the cell membrane was quantified. The cells were divided into regions based on the relative LAMP1 intensity as follows: region 0 (0–20%), region 1 (20–40%), region 2 (40–60%), region 3 (60–80%), and region 4 (80–100%). i LAMP1 fluorescence intensities in each region of MDA-MB-231 cells. The intensity of region 0 was excluded. Points and connecting lines, means; bars, SEMs. j LAMP1 fluorescence intensity in the cell periphery (regions 3 and 4). More than 20 cells per group were assessed in three independent experiments, and the results are shown as a scatter plot. Bars, SEMs. k Matrigel chemoinvasion assay of MCF-7 cells with or without IR treatment. l Immunofluorescence images of LAMP1 in MCF-7 cells. Green, LAMP1; red, α-tubulin; blue, DAPI. Bar, 10 μm. m LAMP1 fluorescence intensities in each region of MCF-7 cells. n LAMP1 fluorescence intensity in the cell periphery (regions 3 and 4) of MCF-7 cells. Bars, SEMs. All column graphs show the means with SEMs of three (d, e, k) or four (a–c) independent experiments. The results of each group in column graphs were normalized against that of the untreated group. *P < 0.05; **P < 0.01; ***P < 0.001; ns, not significant.