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. 2021 Mar 22;11:6489. doi: 10.1038/s41598-021-85833-y

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Loading efficiency of GF-EVs with HSP70 protein. (A) Flowchart summarizing the procedure of GF-EV loading with HSP70. (B) Cryo-EM images of GF-EVs loaded with HSP70. White arrows indicate vesicles with intact membrane, black arrows indicate vesicles with broken membrane. Scale bars are 50 nm. Inset—size distribution histogram. A total of 157 particles was analyzed. (C) Fluorescence of Alexa Flour 647 labeled HSP70 in the initial mixture and loaded samples of GF-EVs, as well as washing filtrates F1 and F10. N = 3. Data are presented as means ± standard deviation (SD). *, p < 0.05; ***, p < 0.001 as estimated by one-way ANOVA followed by multiple comparisons Tukey’s post-hoc analysis. (D) Western blot of HSP70 in the initial and loaded samples of GF-EVs. The full-length blot image used to generate panel is shown in Supplementary Fig. S2.