GSM8125281: PA+rapamycin exosomes1; Homo sapiens; miRNA-Seq
Conditioned media was collected and centrifuged serially at 3000 × g for 20 min and 10,000 × g for 30 min at 4 °C.  Then, the supernatants were passed through a 0.22-μm pore polyethersulfone membrane filter (Millipore) and ultracentrifuged at 100,000 × g (Optima XPN-100, Beckman, USA) for 2 h to isolate pelleted exosomes. The pellets were resuspended in PBS and ultracentrifuged at 100,000 × g for 2 h at 4 °C. The isolated exosomes were suspended in PBS for characterization and the following experiments.Total RNA extracted from purified exosomes was prepared, and the RNA total amount and fragment distribution were detected before the establishment of the sequencing library for each sample using the Small RNA Sample Pre Kit. A total amount of 3 μg total RNA per sample was used as input material for the small RNA library. Sequencing libraries were generated using NEBNext®Multiplex Small RNA Library Prep Set for Illumina®(NEB, USA.)following manufacturer's recommendations and index codes were added to attribute sequences to each sample.