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. 2022 Apr 6;13:1875. doi: 10.1038/s41467-022-29481-4

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© The Author(s) 2022

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 Time-lapse images of wild-type (WT) or SMS-KO HeLa cells expressing GFP-tagged EqtSM or EqtSol and treated with 1 mM LLOMe for the indicated time. b Time-course plotting Eqt-positive puncta per 100 μm² cell area in cells treated as in (a). Data are means ± SD. n = 9 cells for WT + EqtSM, 9 cells for WT + EqtSol, 8 cells for SMS-KO + EqtSM over two independent experiments. c Time-lapse images of wild-type HeLa cells co-expressing GFP-tagged EqtSM (green) and mCherry-tagged Gal3 (magenta) treated with 1 mM LLOMe for the indicated time. d Time-course plotting Eqt- and Gal3-positive puncta per 100 μm² cell area in cells treated as in (c). Data are means ± SD. n = 17 cells. e RAW264.7 cells expressing GFP-tagged EqtSM or EqtSol (green) were infected with mCherry-expressing wild-type (WT) or translocation-defective (ΔRD1) mutant strains of Mycobacteria marinum (magenta). Live-cell fluorescence micrographs were captured 2 h post-infection. f Percentage of cells treated as in (e) showing co-localization of mCherry-expressing M. marinum and EqtSM-positive puncta. n = 30 cells per condition over two independent experiments. g Time-lapse images of wild-type (WT) or SMS-KO HeLa cells expressing GFP-tagged EqtSM or EqtSol and locally wounded by a brief pulse from a 2-photon laser. h Time-course plotting Eqt-associated fluorescence at the laser-induced wound site in cells treated as in (g). Data are means ± SD. n = 7 cells for WT + EqtSM, 4 cells for WT + EqtSol, 6 cells for SMS-KO + EqtSM over two independent experiments. Scale bar, 10 µm.