GSM9112037: LPL:VLDL, Rep5; Homo sapiens; ncRNA-Seq
Following 18 hours of incubation without or with HP, in the absence or presence of NOX inhibitors, media were removed and the cells were washed with 1X PBS, then 1 mL of TRIzol® Reagent (cat. #15596018, Invitrogen, Thermo Fisher Scientific) was pipetted into each well. RNA was extracted as described by the manufacturer. Isolated RNA was resuspended in 40 μL of nuclease-free water. Genomic DNA contamination was removed by treating the isolated RNA with the TURBO DNA-free™ Kit (cat. #AM1907, Invitrogen, Thermo Fisher Scientific) per the manufacturer's instructions. The RNA concentration and purity were measured after DNase treatment. The RNA quality was examined using a 1% agarose gel electrophoresis. Small RNA libraries were prepared at The Centre for Applied Genomics (Toronto, ON, Canada) using the NEBNext® Multiplex Small RNA Library Prep Kit for Illumina® (Index Primers 1-48) (cat. #E7560, New England Biolabs, Whitby, ON, Canada) following the manufacturer's protocol. Pooled libraries of equimolar quantities were sequenced on the Illumina NovaSeq X Series platform using a partial lane of a 25B flow cell.