GSM7949395: IR-4; Mus musculus; RNA-Seq
Cell supernatants were centrifuged at 10,000 ×g for 45 min at 4 °C to remove the larger vesicles. The supernatants were extracted and filtered through a 0.45 µm filter. Then, the samples were centrifuged again at 10,000 ×g and at 4 °C for 70 min and resuspended in PBS. RNA high throughput sequencing was performed by Cloud-Seq Biotech (Shanghai, China). Briefly, total RNA was used for removing the rRNAs with GenSeq® rRNA Removal Kit (GenSeq, Inc.). Then, the rRNA-depleted samples were subjected to library construction with GenSeq® Low Input RNA Library Prep Kit (GenSeq, Inc.) according to the manufacturer's instructions. Libraries were controlled for quality and quantified using the BioAnalyzer 2100 system (Agilent Technologies, Inc., USA). Library sequencing was performed on an Illumina Novaseq instrument with 150bp paired end reads.