medaka larvae(within 24 hours post-hatch) were exposed to 0(control),0.2nM,2nM and 20nM of HBCD(dimethyl sulfoxide with a final concentration of 1:30000 v/v water )for 7 days.In all experiments,90% of the contaminated water was changed every day for the duration of the expose period.Each HBCD treatment had three replicates with 100 larvae for each Petri dish
Growth protocol
Fish was maintained in the laboratory in aerated 30‰ filtered seawater.Medaka was The brood stock was kept under a 14: 10 h light: dark photo-period and the dissolved oxygen level at 5-7mg/L，temperatures of 28±2℃.
Extracted molecule
total RNA
Extraction protocol
At the end of the treatment period,30 larvae/sample were pooled, homogenized in RNA iso Plus(Takara Biotechnology Co., Ltd) and stored in -80 until RNA extraction.Total RNA was amplified and labeled by Low Input Quick Amp Labeling Kit, One-Color (Cat#5190-2305, Agilent technologies, Santa Clara, CA, US), followed the manufacturer’s instructions. Labeled cRNA were purified by RNeasy mini kit (Cat#74106, QIAGEN, GmBH, Germany).
Label
Cy3
Label protocol
Each Slide was hybridized with Cy3-labeled cRNA using Gene Expression Hybridization Kit (Agilent technologies, Santa Clara, CA, US) in Hybridization Oven
Hybridization protocol
After 17 hours hybridization, slides were washed in staining dishes (Cat#121, Thermo Shandon, Waltham, MA, US) with Gene Expression Wash Buffer Kit(Cat#5188-5327, Agilent technologies, Santa Clara, CA, US), followed the manufacturer’s instructions。
Scan protocol
Slides were scanned by Agilent Microarray Scanner (Cat#G2565CA, Agilent technologies, Santa Clara, CA, US) with default settings，Dye channel: Green ,Scan resolution=5μm, PMT 100%，10% ，16bit.
Description
medaka larvae,7days,High_20,replicate 2
Data processing
Feature Extraction software 10.7 (Agilent technologies, Santa Clara, CA, US) Raw data were normalized by Quantile algorithm, Gene Spring Software 11.0 (Agilent technologies, Santa Clara, CA, US).
