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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. 6 — a Western blotting analyses of whole-cell lysates (WCL) from the indicated NCI-H1975 cells stably expressing shNC, shRAB31 or shFLOT1 and shFLOT2. b Western blotting analyses of the concentrated conditional media from the indicated stable NCI-H1975 cells used in a under serum starvation (SS). c Immunofluorescence of endogenous EGFR (green) and CD63 (red) in the indicated stable NCI-H1975 cells used in a under SS. d Western blotting analyses of the concentrated conditional media from the indicated stable NCI-H1975 cells under serum starvation (SS). e Western blotting analyses of the concentrated conditional media from the indicated stable HeLa cells under serum starvation (SS). f Western blotting analyses of WCL and immunoprecipitates (IP) from NCI-H1975 cells treated with afatinib or erlotinib under SS. g Western blotting analyses of the concentrated conditional media from the indicated stable NCI-H1975 cells under SS. h Representative clone images of PC9-GFP cells treated with the concentrated conditional media derived from the indicated stable NCI-H1975 cells without or with erlotinib. i Quantification of the numbers of each clone for h. Data are means ± SD of cell numbers in each clone with PBS (n = 75), Vector (n = 82), RAB31^WT (n = 62), RAB31^Q65L (n = 62), RAB31^R77Q (n = 65) or RAB31^3YF (n = 84). j Representative clone images of PC9-GFP cells treated with the pure small EV (sEV) derived from the indicated stable NCI-H1975 cells without or with erlotinib. k Quantification of the numbers of each clone for j. Data are means ± SD of cell numbers in each clone with Vector (n = 71), RAB31^WT (n = 80) or RAB31^3YF (n = 80). Unpaired t-test was used to analyze the difference between the two groups. ****P < 0.0001, NS, no statistical significance. Scale bars, 10 μm (c) and 100 μm (h and j).