meRIP-seq of villous tissue
Total RNA was extracted using Trizol reagent (Takara, Dalian, China). Agilent 2100 Bioanalyzer (Agilent, CA, USA) and NanoDrop 2000 (Thermo Fisher, MA, USA) was used to analyze the total RNA quality and quantity.The total cell RNA is fragmented into ~100-nt-long oligonucleotides using fragmentation buffer (part # CS220011) under elevated temperature. Then validation of post-fragmentation size distribution by Agilent 2100 Bioanalyzer with an Agilent RNA 6000 Kit. The Magna ChIP Protein A/G Magnetic Beads were subjected to incubate for 30 minutes at room temperature with m6A-specific antibody (Part # MABE1006) in IP buffer. The mixture was then incubated with MeRIP reaction mixture for 2 hours at 4C. Then eluted RNA and analyzed MeRIPed RNA by deep sequencing on an Illumina Novaseq 6000 platform by Gene Denovo Biotechnology Co. (Guangzhou, China). following the vendor's recommended protocol.