RNA-seq raw data in primary thyroid cells (negative control)
The total RNA samples for RNA-seq were extracted by TRIzol reagent (Invitrogen) and then treated byDNase-1 (Ambion) in order to eliminate the DNA contamination. RNA concentration was determined by using Qubit 2.0 Fluorometer (Agilent Technologies) with an RNA HS Assay Kit. The integrity of the RNA samples were assessed by BioAnalyzer (Agilent). All the RNA Integrity Numbers (RIN) were greater than 8. The purified RNAs with no visible sign of genomic DNA contamination from the HS Nanochip tracings were used for total RNA library generation. Furthermore, Illumina TruSeq Stranded Total RNA Sample Prep Kit with Ribo-Zero Gold (catalog #RS-122-2201) was used to transform RNA into cDNA after removing rRNA and mitochondrial RNA. The RNA-seq libraries were prepared according to the manufacturer’s protocol. Finally, 75bp paired-end sequencing was performed using the Illumina HiSeq 2500 system.