Total RNA was isolated from cells/tissues using the Magzol Reagent (Magen, China)according to the manufacturer's protocol, The quantity and integrity of RNA yield was assessed by using the K5500(BeijingKaiao, China) and the Agilent 2200 TapeStation (Agilent Technologies, USA) separately. Briefly, rRNAs were removed from Total RNA using RiboCoprRNA Depletion Trial kit(LEXOGEN， Australia) , thenRNA was treated with RNase R (Epicentre, USA) and fragmented to approximately 200bp. Subsequently, thepurified RNA fragments were subjected to first strand and second strand cDNA synthesis following by adaptorligation and enrichment with a low-cycle according to instructions of NEBNext® Ultra™ RNA Library Prep Kitfor Illumina (NEB, USA). The library products were evaluated using the Agilent 2200 TapeStation andQubit(Thermo Fisher Scientific, USA) and then sequenced on Illumina (Illumina, USA) withpaired-end 150bp at Ribobio Co. Ltd (Ribobio, China). circRNA-seq