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. 2023 Dec 18;14:8398. doi: 10.1038/s41467-023-44134-w

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© The Author(s) 2023

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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Fig. 2 — a Schematic for MV with high ACE2 expression (aMV) preparation and its mechanism of entrapping the SPV. b Confocal laser scanning microscopy (CLSM) images showing the ACE2 protein expression on wild-type 293 T cells transfected with control empty plasmid (WT-293T) or ACE2-plasmid (ACE2-293T). White: ACE2 antibody labeled ACE2 protein. Blue: 4′,6-diamidino-2-phenylindole (DAPI) labeled cell nuclei. c CLSM images of ACE2-293T cells after cytochalasin B treatment. Red: carboxyfluorescein succinimidyl ester labeled cytoplasm (CSFE). d CLSM and transmission electron microscope (TEM) images of aMV. Red: 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindodicarbocyanine,4-chlorobenzenesulfonate salt (DiD) labeled cell membrane. e CLSM images of ACE2 protein expression on vesicles that were obtained from WT-293T cells (nMV) and aMV. Red: DiD labeled cell membrane. White: ACE2 antibody labeled ACE2 protein. f Flow cytometry analysis of ACE2 protein expression on nMV and aMV. The ACE2 protein were labeled with the ACE2 antibody. g Stimulated emission depletion (STED) microscopy image of SPV (wild-type) internalized within aMV. Green: fluorescein isothiocyanate (FITC) labeled SPV. Red: DiD labeled cell membrane. h TEM images of ultrathin section of aMV after 24 h of challenge with SPV (wild-type). The green arrow indicated SPV. In the enlarged image (right), SPV was marked with false color (green). i CLSM images of ACE2-293T cells that were treated with the indicated agents and then challenged with SPV (wild-type), accompanied with the corresponding quantification of relative infection rate. Green: GFP was expressed in infected cells. Blue: DAPI labeled cell nuclei. j Relative infection rates of ACE2-293T cells that were treated with PBS or aMV and then challenged with the indicated SPVs with S protein mutation. Quantitative data in f, i, and j represent as means ± S.E.M., n = 3 biologically independent experiments. Statistical significance was calculated using two-tailed unpaired t-test (f, j) and one-way ANOVA with multiple comparison tests (i). All P-values are indicated. Source data are provided in the Source data file.