HuR targetome analysis in THP-1 macrophages using HuR FLASH-Seq.
FLASH, a novel method, was utilized to identify binding sites of RNA-binding proteins (RBPs) with single-nucleotide resolution (Nucleic Acids Res 2020;48:e15). In order to identify HuR target genes in THP-1 macrophages after circARCN1 regulation, we performed FLASH experiments using the anti-HuR antibody on THP-1 macrophages with circARCN1 knockdown or overexpression. The resulting FLASH products were then subjected to RNA sequencing. Overall design: HuR FLASH experiments were conducted using the anti-HuR antibody on wild type THP-1 macrophages (two replicates, THP1-FLASH-1 and THP1-FLASH-2), and THP-1 macrophages with circARCN1 knockdown (THP1-SH-FLASH) or overexpression (THP1-OE-FLASH). The resulting FLASH products were then subjected to RNA sequencing.