RNA immunoprecipitation (RIP) sequencing

RIP-RNA high-throughput sequencing was performed by Shanghai Cloud-seq Biotech Co.,Ltd. RIP was performed as previously described ⁴³. RNA was extracted using TRIzol^® and ribosomal RNAs (rRNAs) were removed using NEBNext rRNA Depletion Kit (New England BioLabs, Inc.). RNA libraries were constructed using rRNA-depleted RNAs with NEBNext^® Ultra™ II Directional RNA Library Prep Kit (New England BioLabs, Inc.) according to the manufacturer's instructions. Libraries were controlled for quality and quantified using the BioAnalyzer 2100 system (Agilent Technologies, Inc.). Library sequencing was performed on an Illumina HiSeq instrument with 150-bp paired end reads.