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. 2021 Jul 23;9:698639. doi: 10.3389/fcell.2021.698639

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Copyright © 2021 Cochran and Kornbluth.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Orientation of transmembrane proteins in NK3.3-derived EVs. (A) Twenty micrograms of NK3.3 EVs were treated with either PBS (-), Pronase (+), or Pronase + saponin (+s). Samples were then boiled in Laemmli buffer; immunoblot analysis was performed using antibodies to protein epitopes that have a vesicle membrane, cytoplasmic-facing orientation or contained within the EV lumen. (B) Conceptualization of NK3.3 EV composition depicting the localization of proteins in NK3.3-derived EVs. The majority of NKLAM and LAMP-1 proteins are positioned in this orientation.