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. 2021 Apr 24;62(3):401–413. doi: 10.1093/jrr/rrab018

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© The Author(s) 2021. Published by Oxford University Press on behalf of The Japanese Radiation Research Society and Japanese Society for Radiation Oncology.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Fig. 1. — Schematic representation of the experimental workflow used in the study. CAFs were isolated from fresh NSCLC specimens and grown in exosome-free culture media. CAF cultures were submitted to single high-dose (1 × 18 Gy) or fractionated (3 × 6 Gy) radiation regimens. CAF-EVs were isolated by sequential centrifugations and ultracentrifugations of conditioned medium from irradiated and non-irradiated CAFs. Purified EVs were resuspended and characterized as follows: CAF-EVs morphology analyzed by immuno-gold labeling for EV-markers and imaged by TEM and SIM; protein expression of specific EV-markers quantified by Western blotting and total CAF-EVs was quantified by flow cytometry. The CAF-EV protein cargo was analyzed by LC-MS/MS. Scale Bar = 15 μm.