tissue: Bone marrow gender: male age: 6-8week strain: C57BL/6
Treatment protocol
The mice were pretreated with 10mg/kg Amifostime (i.p. 0.5 hr pre-IR), 0.2mg/kg CBLB502 (i.p. 0.5 hr pre-IR) and 150mg/kg Nilestriol (i.g. 48 hr pre-IR), and then radiated with 8 Gy gamma-radiation. At 12 hr post-radiation, samples of the bone marrow or hyprothalamus (pretreated with PBS or 10 mg/kg Amifostine) were obtained from the three mice per group.
Extracted molecule
total RNA
Extraction protocol
Total RNA was extracted, pooled, and purified using miRNeasy Mini Kit (QIAGEN, GmBH, Germany) following the manufacturer’s instructions, and the RNA integrity number was verified on an Agilent Bioanalyzer 2100 system (Agilent Technologies, Santa Clara, CA, USA).
Label
Cy3
Label protocol
Total RNA was labeled by miRNA Complete Labeling and Hyb Kit (Cat#5190‐0456, Agilent technologies, Santa Clara, CA, US) followed the manufacturer’s instructions, labeling section.
Hybridization protocol
Each slide was hybridized with 100ng Cy3‐labeled RNA using miRNA Complete Labeling and Hyb Kit (Cat# 5190-0456, Agilent technologies, Santa Clara, CA, US) in hybridization Oven (Cat#G2545A, Agilent technologies, Santa Clara, CA, US) at 55℃，20rpm for 20 hours according to the manufacturer’s instructions, hybridization section. After 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).
Scan protocol
Slides were scanned by Agilent Microarray Scanner (Cat#G2565BA, Agilent technologies, Santa Clara, CA, US) and Feature Extraction software 10.7 (Agilent technologies, Santa Clara, CA, US) with default settings.
Description
miRNA expression after 12hr in 0 Gy-irradiated mice bone marrow pretreated with PBS (i.p.)
Data processing
Raw data were normalized by Quantile algorithm, Gene Spring Software 11.0 (Agilent technologies, Santa Clara, CA, US).
