tissue: mammary gland strain: Wistar Han treatment: none
Treatment protocol
Females and males Wistar Han rats at eight weeks of age (Harlan France Sarl, Gannat, France) were maintained in an animal facility and fed a purified diet (phytoestrogen-free, INRA, France) and water ad libitum. Procedures for handling and experimentation followed ethical guidelines and were previously described [37, Reprod Toxicol]. At gestational day 1, determined by the presence of intravaginal sperm plug, the dams were randomly divided into groups of 15. From the first gestational day (GD1) until the weaning (PND 21), each pregnant female were orally gavaged once daily with genistein (G), vinclozoline (V) alone or in combination (GV) at 1 mg/kg body weight/day, as previously described ( ). At weaning, immature rats were dispatched. Female offspring were randomized (one female per litter), identified by implanted chips, and fed the purified diet until sacrifice (PND 35 and PND 50) (Fig. 1). Twenty pubertal or cycling female offspring animals per treatment group were used for subsequent analysis.
Growth protocol
Females and males Wistar Han rats at eight weeks of age (Harlan France Sarl, Gannat, France) were maintained in an animal facility and fed a purified diet (phytoestrogen-free, INRA, France) and water ad libitum. Procedures for handling and experimentation followed ethical guidelines. At gestational day 1, determined by the presence of intravaginal sperm plug, the dams were randomly divided into groups of 15. From the first gestational day (GD1) until the weaning (PND 21), each pregnant female were orally gavaged once daily with genistein (G), vinclozoline (V) alone or in combination (GV) at 1 mg/kg body weight/day, as previously described. At weaning, immature rats were dispatched. Female offspring were randomized (one female per litter), identified by implanted chips, and fed the purified diet until sacrifice (PND 35 and PND 50). Twenty pubertal or cycling female offspring animals per treatment group were used for subsequent analysis
Extracted molecule
total RNA
Extraction protocol
Total RNA was extracted from individual mammary glands with Trizol extraction.
Label
biotin
Label protocol
cDNA is end labelled with biotin using Terminal Transferase (using the WT terminal labelling kit of Affymetrix)
Hybridization protocol
cDNA is then hybridized to GeneChip® rat Gene (Affymetrix) at 45°C for 17 hours
Scan protocol
After overnight hybridization, chips are washed on the fluidic station FS450 following specific protocols (Affymetrix) and scanned using the GCS3000 7G
Data processing
Raw data were preprocessing using the Robust Multichip Algorithm
