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. 2020 Sep 21;31(2):157–177. doi: 10.1038/s41422-020-00409-1

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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. 4 — a, b Western blotting analyses of whole-cell lysates (WCLs) and immunoprecipitates (IP) at their endogenous levels from NCI-H1975 cells using anti-FLOT1 antibody (a) or anti-FLOT2 antibody (b). c, d Western blotting analyses of WCL and IP from HEK-293T cells co-expressing the indicated plasmids. e Immunofluorescence of erlin1-HA, erlin2-HA, prohibitin1-HA or prohibitin2-HA (green) with Flag-RAB31^Q65L (red) in HeLa cells stably expression Flag-RAB31^Q65L and transiently expressing the indicated plasmids under serum starvation (SS). f Immunofluorescence of stomatin-HA or STOML3-HA (red) and Flag-RAB31^Q65L (magenta) with CD63-GFP (green) in HeLa cells stably expression Flag-RAB31^Q65L and transiently expressing the indicated plasmids under SS. g Immunofluorescence of FLOT1-HA, FLOT2-HA or stomatin-HA and Flag-RAB31^Q65L (magenta) with CD63-GFP (green) in the indicated stable HeLa cells transiently expressing the indicated plasmids under SS. h Immunofluorescence of FLOT1-SPFH-HA, FLOT2-SPFH-HA, FLOT1-flotillin-HA, or FLOT2-flotillin-HA (red) and Flag-RAB31^Q65L (magenta) with CD63-GFP (green) in the indicated stable HeLa cells transiently expressing the indicated plasmids under SS. i Immunofluorescence of Sto-flotillin1-HA (red) chimeras and Flag-RAB31^Q65L (magenta) with CD63-GFP (green) in the indicated stable HeLa cells transiently expressing the indicated plasmids under SS. j Western blotting analyses of the concentrated conditional media from the indicated stable HeLa cells stably re-introduced with the indicated plasmids. Scale bars, 10 μm.