GSM4849595: Vehicle_STS-26T-luc (G434_SL01); Homo sapiens; RNA-Seq
Total RNA was extracted from the cells (MPNST or Schwann cells) with or without drug or vehicle-treated cells using the RNeasy Mini Kit (QIAGEN) according to the manufacturer's instructions. The NanoDrop 2000 spectrophotometer (Thermo Scientific) and 2100 Bioanalyzer (Agilent) were used to assess RNA quantity and quality, with the calculation of RNA integrity number (RIN). Samples were included for RNA-Seq if RIN > 6 and total RNA> 500 ng. Illumina RNA-Seq was performed according to the manufacturer's instructions. Briefly, the cDNA libraries were prepared using TruSeq® Stranded mRNA Library Prep kit as per the manufacturer's instructions. The final product was assessed for its size distribution and concentration using the BioAnalyzer High Sensitivity DNA Kit (Agilent). Pooled libraries were diluted to 3 nM using 10 mM Tris-HCl, pH 8.5 and then denatured using the Illumina protocol. The denatured libraries were loaded onto an S1 flow cell on an Illumina NovaSeq 6000 (Illumina) and run for 2X50 cycles according to the manufacturer's instructions.