Zebrafish whole embryo co-treated with IOP + T3, replicate 2
Zebrafish (Danio rerio) 3hpf old embryos were placed in to a 24 wells plate with 20 embryos per well and 2 mL of exposure solution : 0.6 mg /L IOP and 0.8 g/L T3 . They were exposed in light-dark cycles (12h-14h), at 21C and medium were renewed every day for 4 days. Tissue  fractionation was done with tissue lyser II (QIAGEN) and a stainless-steel ball (INOX AISI 304 grade 100 AFBMA) in 500 L RNAble (Eurobio). Tubes were transferred on ice and 100 L chloroform added. Samples were then centrifugated 12 000 g at 4C during 15 min. Next, 175 L of AquaPhenol (Q-Biogene: AquaPH01) and 100 L of chloroform were added to 350 L of supernatant. Following centrifugation 12 000 g at 4C during 15 min, RNAs from 250 L of the supernatant were recovered with RNeasy Mini Kit (Qiagen: 74104). Sequencing libraries were prepared with TruSeq RNA Sample Prep v2 kit.