RNASeq of Xenopus whole tadpole treated with T3
Animal treatments were carried out according to the OECD XETA test guideline. Xenopus laevis THb/ZIP-GFP transgenic line tadpoles at NF stage 45 were placed in to a 6 well plate with 10 tadpoles per well and 8 mL of exposure solution, 3.25 g.L-1 T3. Tadpoles were exposed in the dark, at 21C and medium were renewed every dayfor 3 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.