RNA from cells, exosomes, and plasma was isolated using a QIAzol Lysis reagent and total RNA was isolated using Direct-zol™ RNA MiniPrep isolation kit (Zymo Research). 100 μL of exosome suspension or plasma from plasma of patients with HNSCC or healthy subjects were mixed with 300 μL QIAzol lysis buffer, and the mixture was processed according to the standard protocol. Quantity and quality of the RNA were determined by NanoDrop 1000 (260/280 and 260/230 ratios).

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