Cellular and intravesicular miRNAs were extracted from centrifugation pellets or immunoprecipitates. These were lysed with 700 µL of Qiazol (Qiagen, Venlo, The Netherlands). Extraction was then performed using the “miRNEasy mini kit” (Qiagen) according to the supplier’s recommendations. Eluates were stored at −80 °C before analysis.
The miRNAs from culture supernatants, centrifugation supernatants or filtrates were extracted from 200 µL of the sample. The extraction was then performed using the miRNEasy Advanced Serum/Plasma Kit (Qiagen) according to the supplier’s recommendations.
Extractions from the different kits were performed on the Qiacube automated system (Qiagen).
Cel-miR-39-3p spike-in control, obtained from Qiagen, was added to the samples to either check the quality of the purification or normalize the bkv-miR-B1-5p qPCR results after successive filtrations and or centrifugations.
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