RNA-Seq of HCAEC treated with siRNA against scrambled or VKORC1 or VKORC1L1
Human coronary artery endothelial cells (HCAEC) were cultured under standard conditions and harvested at 24h point following siRNA transfection. Cells were washed with PBS and collected directly into lysis reagent (Trizol)for RNA isolation. Total RNA was extracted using TRIzolâ€“chloroform phase separation followed by isopropanol precipitation and ethanol washes. RNA was resuspended in RNase-free water and assessed for concentration, purity, and integrity prior to library preparation. RNA-seq libraries were prepared from total RNA using poly(A) mRNA enrichment and strand-specific library preparation. mRNA was fragmented, reverse transcribed, end-repaired, adapter-ligated, PCR-amplified, and quality-checked before sequencing.