Exosome labeling and uptake

Huaiming Sang; Weifeng Zhang; Lei Peng; Shuchun Wei; Xudong Zhu; Keting Huang; Jiajia Yang; Meihong Chen; Yini Dang; Guoxin Zhang

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Exosome labeling and uptake

HS Huaiming Sang

WZ Weifeng Zhang

LP Lei Peng

SW Shuchun Wei

XZ Xudong Zhu

KH Keting Huang

JY Jiajia Yang

MC Meihong Chen

YD Yini Dang

GZ Guoxin Zhang

This method is extracted from research article: Cell Death Dis, Jan 2022

Exosomal circRELL1 serves as a miR-637 sponge to modulate gastric cancer progression via regulating autophagy activation

DOI: 10.1038/s41419-021-04364-6

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Exosomes were labeled with PKH26 red fluorescent dye (Sigma-Aldrich, USA) following the manufacturer’s protocol. A total of 50 μg of exosomes were resuspended in 100 μl PBS and were added to 5 × 10⁵ gastric tumor cells. Faction was stained with phalloidin-FITC (green), and DAPI (blue) was used to keep the nucleus. To track exosomes, exosomes secreted from GC cells were labeled with PKH26 red fluorescent dye (Sigma-Aldrich, USA), while circRELL1 was marked with GFP (green), and DAPI (blue) was used to mark the nucleus. Subsequently, a fluorescence microscope (Zeiss, LSM700B, Germany) was adopted to examine stained cells. The uptake capacity of AGS and SGC-7901 into exosomes (exos-Vector/exos-circRELL1) was assessed via immunofluorescence assays and qRT-PCR.

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