RIP-seq analysis of HUVEC cells overexpressed wild-type or mutanted DDX24

To investigate the impact of DDX24 mutation on the RNA binding activity in HUVEC cells, we conducted RIP-seq analysis. Overall design: For RIP-seq, RNA interacting with DDX24-WT or DDX24-?RBD was isolated following the RIP protocol. The immunoprecipitated RNA was then subjected to mRNA selection using Oligo dT-based Poly(A) mRNA Magnetic Isolation Module (NEB) and prepared for sequencing libraries using Ultra II Directional RNA Library Prep Kit for Illumina (NEB). Libraries were pooled and sequenced on an Illumina Hiseq 3000 PE150 platform.