RNA-seq for NKX2-3 knockout tumor pericytes

This study aims to investigate genes regulated by NKX2-3 For NKX2-3-knockdown experiments, TPCs derived from CRC patients with liver metastasis were infected with Guide RNA of NKX2-3 for 48 h and selected by puromycin (2 mg/mL). Total RNA was extracted using TRIzol reagent (Life technologies) according to the manufacturer manual. The RNA concentration and integrity were evaluated by NanoDrop 2000. Then, the samples were inspected by agarose gel electrophoresis, Nanodrop and Agilent 2100. Then the fragmented RNA pieces were used to construct library by Hieff NGS® Ultima Dual-mode mRNA Library Prep Kit and sequenced with Illumina Novaseq™ 6000