GSM5769483
HeLa cells were ground into fine powder to extract RNA. Total RNA was extracted with TRIzol (Ambion). RNA was purified with two phenol-chloroform treatments, and then the DNA was digested with RQ1 DNase (Promega, Madison, WI, USA). SmartSpec Plus (Bio-Rad, USA) was used to redetermine the quality and quantity of purified RNA by measuring the absorbance at 260 nm/280 nm (A260/A280). The integrity of RNA was verified by 1.5% agarose gel electrophoresis. One microgram of the total RNA from each sample was used for RNA-seq library preparation with the VAHTS Stranded mRNA-seq Library Prep Kit (Vazyme). The polyadenylated mRNAs were converted into double-stranded cDNA after purification and fragmentation. The DNAs were ligated to VAHTS RNA Adapters (Vazyme) after end repair and A tailing. Heat-labile UDG was used to digest the purified ligation products corresponding to 200-500 bp, after which the fragments were amplified and purified. Single-stranded cDNA was quantified, stored at -80°C and sequenced.  The libraries were prepared following the manufacturer's instructions. The Illumina HiSeq X Ten system was used for 150 nt paired-end sequencing, and high-throughput sequencing was performed. For high-throughput sequencing, the libraries were prepared following the manufacturer's instructions and applied to the Illumina HiSeq X Ten system for 150 nt paired-end sequencing.