GSM7765085: H9 cells, METTL3_Input; Homo sapiens; RIP-Seq
RIP assay was performed on METTL3-SMASh tagged hNPCs by SeqHealth (Wu-han, China). The cells were treated with cell lysis buffer. The 10% lysis sample was stored and named “input”, and 80% was used in immunoprecipitation reactions with METTL3 antibody（Abcam, ab195352, 10 μg / IP）and named “IP”, and 10% was in-cubated with rabbit IgG (Cell Signaling Technology) as a negative control and named “IgG”, respectively. The RNA of input and IP was extracted using TRIzol reagent (Invitrogen, 15596026). The stranded RNA sequencing library was constructed by using KC-DigitalTM Stranded mRNA Library Prep Kit for Illumina (Wuhan Seqhealth, DR08502) following the man-ufacturer's instruction. The kit eliminated duplication bias in PCR and sequencing steps, by using unique molecular identifier (UMI) of 8 random bases to label the pre-amplified cDNA molecules. The library products corresponding to 200-500 bps were enriched, quantified and finally sequenced on Novaseq 6000 sequencer (Illumina) with PE150 model.