Illumina HiSeq 2500 paired end sequencing; RNA-seq of immortalized rat thyrocytes (PCCl3) treated with two thyroid disrupting chemicals (ethylenethiourea and chlorpyrifos) against untreated controls
RNA-seq of immortalized rat thyrocytes (PCCl3) treated with two thyroid disrupting chemicals (ethylenethiourea and chlorpyrifos) against untreated controls
PCCl3 were maintained in Coon’s modified F12 medium (EuroClone) supplemented with 5% newborn bovine serum (HyClone Laboratories) and six hormones 16. Cells were exposed for 7 days to CPF (6x10-7 M, 6x10-8 M, 6x10-9 M, Greyhound Chromatography F2057) and ETU (6x10-8 M, 6x10-9 M, 6x10-10 M, 2-Imidazolidinethione, Sigma-Aldrich I504) and their combinations. Total RNA was isolated from treated and control cells using Trizol reagent (Invitrogen) following manufacturer’s instructions. Total RNA samples were treated with DNasi (NEB), purified with RNasy Mini Kit (Qiagen) and controlled with Agilent 2100 bioanalyzer (Agilent Technologies). RNAs, 3 µg/each obtained from three biological replicates, were pooled to minimize sample-to-sample variations. 1μg of each RNA pool was used for library preparation with TruSeq Stranded total RNA Sample Prep Kit (Illumina Inc.) according to the manufacturer’s instructions.
Experimental Factor: chlorpyrifos and ethylenethiourea
compound
Experimental Factor: 50 and 0.6
dose