Illumina NovaSeq 6000 paired end sequencing; NFKB1 and NFKB2 differently modulate transcriptional profile of MT-2 cells
NFKB1 and NFKB2 differently modulate transcriptional profile of MT-2 cells
Jurkat and MT-2 cells were provided by the Japan National Institute of Biomedical Innovation, Health and Nutrition, Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan). The cell lines were cultured in Roswell Park Memorial Institute medium (Life Technologies) supplemented with 10% fetal bovine serum (Gibco). Mycoplasma testing (MycoAlert Quick Mycoplasma Detection Kit, Lonza) was routinely conducted. Mycoplasma contamination was not detected during the study period.  RNA was extracted from the 1 x106 cells by QIAzol Lysis Reagent (QIAGEN). Concentrations of nucleic acids were measured by nanodrop (all experiments) and/or Qubit HS RNA-kit (only for RNA-seq). RNA quantity and quality were measured by Agilent 2100 Bioanalyzer (Agilent technologies). RNA integration values (RIN) >8 were considered to proceed to the further analysis. RNA-seq libraries were generated using Illumina TruSeq Stranded Total RNA kits. The resultant cDNA libraries were quantified via Qubit and assessed for quality via Bioanalyzer.