RNA-Seq of Blood Vessel
Total RNA was extracted from both stable shFTO VSMCs and their respective control samples using Trizol reagent sourced from ThermoFisher (USA). Subsequently, rRNA was depleted from these RNA samples using the Ribo-Zero rRNA Removal Kits (Illumina, San Diego, CA, USA), adhering strictly to the manufacturer's guidelines. rRNA-depleted RNA was then utilized to construct RNA libraries with the TruSeq Stranded Total RNA Library Prep Kit (Illumina, San Diego, CA, USA), following the manufacturer's instructions meticulously.Quality control and quantification of the constructed libraries were conducted using the BioAnalyzer 2100 system (Agilent Technologies, Inc., USA). These libraries, at a concentration of ten picometers, were processed to convert them into single-stranded DNA molecules, which were then captured onto Illumina flow cells. In situ amplification was performed to form clusters, and finally, the libraries underwent 150 cycles of sequencing on an Illumina HiSeq Sequencer, all according to the manufacturer's protocols. Novogene (Tianjin, China) provided the services for library preparation and high-throughput sequencing