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Measurement of phagocytosis and endosomal escape of PMVs

ML Mei-jia Lin
SL Shuang Li
LY Lu-jun Yang
DY Dan-yan Ye
LX Li-qun Xu
XZ Xin Zhang
PS Ping-nan Sun
CW Chi-ju Wei
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hUCMSCs cells were stained with Calcein-AM (2 μM) for 30 min or MitoTracker-Green (200 nM) for 1 h before being used for PMV generation. VSV-G viral membrane glycoprotein was prepared as described. PMVs were incubated with or without VSV-G for 20 min and then added to HepG2 cell culture in 96-wells. The plate was centrifuged at 1000 rpm for 10 min to accelerate PMV sedimentation. At 12 h, cells were replenished with fresh culture medium. At 24 h, cells were harvested, stained with Hoechst (10 μg/ml) for 30 min, and then transferred to a 35-mm glass-bottom dish for the examination of phagocytosis and endosomal escape of PMVs using confocal microscopy.

Copyright and License information: The Author(s) ©2020
Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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