Early time-responses in estrogen regulated transcriptome in Atlantic cod (Gadus morhua) exposed to ethynylestradiol
Early time-responses in estrogen regulated transcriptome in Atlantic cod (Gadus morhua) exposed to ethynylestradiol
The fish were sampled at 20 h or 48 h post injection. The fish were killed by a blow to the head. Liver samples were rapidly excised, snap frozen in liquid nitrogen and stored at -80°C for further processing. Liver  total RNA extraction was performed from individual fish. A total of 32 individual liver RNA samples from groups of 20 h time point (Control, BPA (40 μmol/kg bw), MXC (40 μmol/kg bw) and EE2 (50 nmol/kg bw), n = 6 per group) and 48 h time point (Control, n = 3, EE2 = 5) were analyzed. RNA extraction was performed from each sample using TRI Reagent (Sigma) according to the manufacturer's protocols. The concentration and quality of total RNA was assessed using a NanoDrop ND-1000 spectrophotometer (NanoDrop Technologies, Wilmington, DE, USA), and the Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA).  RNA sequencing was performed at the Genomics Core Facility at the University of Bergen, as previously described (Yadetie et al., 2018), according to Illumina TruSeq® Stranded mRNA Sample Preparation Guide on Illumina HiSeq 4000 (Illumina, Inc., San Diego, CA, USA).