Dams were randomly distributed among three groups (3 dams per group) and exposed by caudal injection (0.3 ml/kg body weight) on GD 15, PND 1 and 5. Dams of the control group received vehicle: ethanol 95%, Cremophor EL, and sterile water (1:1:8, v/v); dams of exposed group 1 received 0.002 mg/kg body weight (BW) of BDE-47 in vehicle, and dams of exposed group 2 received 0.2 mg/kg body weight (BW) of BDE-47 in vehicle. One female pup per litter was selected randomly and sacrificed by decapitation on PND 10. Total RNA was extracted from total brain samples of these pups and used for microarray analysis.
Growth protocol
We obtainedtimed pregnant Wistar rats (250–300 g; Charles River Laboratories, St. Constant, Que., Canada) on gestational day (GD) 14 and housed them individually in plastic cages with sawdust bedding, under regulated temperature (21 ± 2 ° C), relative humidity (50 ± 10%) and a 12-hour light/dark cycle. Food (Rodent chow 5075; Charles River Laboratories) and water were provided ad libitum. All animals received care in compliance with the Guide to the Care and Use of Experimental Animals from the Canadian Council of Animal Care and the protocol was approved by our institutional animal research ethics review board.
Extracted molecule
total RNA
Extraction protocol
All tissues were snap-frozen in liquid nitrogen, and stored at -80°C until RNA extraction. RNA was extracted with the RNeasy Mini Kit (Qiagen, Maryland, USA) according to the manufacturer’s instructions.
Label
biotin
Label protocol
Biotinylated cRNA were prepared with the Ambion MessageAmp kit for Illumina arrays
Hybridization protocol
Standard Illumina hybridization protocol
Scan protocol
Standard Illumina scanning protocol
Description
individual pup
Data processing
The raw data were pre-processed and normalized with the lumi Bioconductor package
