GSM9315793: Senescent HUVECs, Rep C; Homo sapiens; RNA-Seq
Total RNA was extracted from the cell pellets using the MagMAX™ mirVana™ Total RNA Isolation Kit (Thermo Fisher Scientific, Waltham, MA, USA) following the manufacturer's instructions for ECs. EVs were processed using the Total Exosome RNA and Protein Isolation Kit (Invitrogen), following the manufacturer's recommendations. Dual-indexed mRNA libraries were prepared using the QuantSeq 3' mRNA-Seq V2 Library Prep Kit with UDI (Lexogen GmbH, Vienna, Austria), according to the user guide. According to the manufacturer's recommendations, 250 ng of starting material was used for cellular RNA analysis, while a low-input protocol (<10ng total) was used for EVs. Library quality was evaluated using the QiaXcel Advanced System (Qiagen), while the quantity was measured using a Quantus Fluorometer (Promega Corporation).