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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. 2 — a Arl8b in the lysosomal fraction of MDA-MB-231 cells. Markers of the nucleus (Histone H3), lysosomes (Rab9) and the cytoskeleton (β-actin) were stained with antibodies. The amount of Arl8b in the lysosomal fraction was normalized against LAMP1. b Total Arl8b in whole-cell lysates of MDA-MB-231 cells after IR treatment was detected by immunoblotting. c, d Arl8b-mVenus overexpression was induced by doxycycline (c) and assessed by confocal fluorescence microscopy (d). Dox, doxycycline. Green, Arl8b-mVenus; red, LysoTracker. Bar, 10 μm. e Lysosomal distribution in Arl8b-mVenus-overexpressing MDA-MB-231 cells. Pseudocolor micrographs with lysosomes shown in red (LAMP1), microtubules shown in blue (α-tubulin), and nuclei shown in white (DAPI). Bar, 10 μm. f LAMP1 fluorescence intensities in each region. The intensity of region 0 was excluded. Points and connecting lines, means; bars, SEMs. g LAMP1 fluorescence intensity in the cell periphery (regions 3 and 4). Bars, SEMs. h Matrigel chemoinvasion assay of Arl8b-mVenus-overexpressing MDA-MB-231 cells. Representative images of the results were obtained from the Matrigel invasion assays. i Immunofluorescence images showing essential proteases in Arl8b-positive lysosomes. Blue, proteases; green, Arl8b-mVenus; red, LysoTracker. Bar, 10 μm. All column graphs show the means with SEMs of three independent experiments. The results of each group in column graphs were normalized against that of the untreated group. *P < 0.05; ***P < 0.001; ns, not significant.