GSM8117756
Cells were lysed by the addition of TRIzol reagent (Invitrogen, 15596018). After homogenizing via pipetting, total RNAs was extracted via addition of chloroform, isopropanol precipitation, ethanol washing, and air dry according to manufacturer's protocol. The extracted RNAs were further digested via DNase I (NEB, M0303) at 37°C for 30 min. The reaction mix were purified via RNA Clean & Concentrator kit (Zymo, R1018). Fragment Analyzer (Agilent) was used to analyze the integrity of purified RNAs. Only samples of RQN > 9.0 were subjected to downstream affinity purification. 10 mL purified RNA was set as Input sample. The remained RNAs were subjected to affinity purification, and set as Enrich or Control samples. Input, Enrich, and Control samples from each biological replicate experiment were subjected to library construction by NEBNext Ultra II RNA Library Prep Kit for Illumina (NEB, E7770) with Poly(A) mRNA Magnetic Isolation Module (NEB, E7490) according to manufacturer's instructions.