Quantification of microRNA Levels with qPCR

Levels of selected microRNA were evaluated by qPCR. To remove heparin traces, RNA was treated with heparinase (Sigma) as was described before (41 (link)). For reverse transcription and qPCR microRNA-specific TaqMan Assays, TaqMan MicroRNA Reverse Transcription Kit and TaqMan Universal Master Mix II no UNG (all Thermo Fisher Scientific) were used according to manufacturer's recommendations. For miR-3679-5p measurement, a reverse transcription stem-loop primer and a primer pair for amplification were designed using sRNAPrimerDB online service (42 (link)). In this case, reverse transcription was performed using TaqMan MicroRNA Reverse Transcription Kit (Thermo Fisher Scientific), and real-time PCR was performed using qPCRmix-HS SYBR (Evrogen). TaqMan Assays and designed primer sequences are indicated in Supplementary Table 1.