cell type: Mes cell line genetic backgroud: RbL/L, Trp53L/L
Treatment protocol
no treatment
Growth protocol
RPMI1640+10%FBS
Extracted molecule
total RNA
Extraction protocol
Total RNAs from the Mes and Epi cell lines were extracted by TRIzol reagent (Life Technologies) according to the manufacturer’s instruction.
Label
Cy3
Label protocol
Total RNAs were amplified and labeled by Low Input Quick Amp Labeling Kit, One-Color (Agilent technologies, Santa Clara, CA, US), following the manufacturer’s instructions.
Hybridization protocol
Each slide was hybridized with 1.65mg Cy3-labeled cRNA using Gene Expression Kit (Agilent technologies, Santa Clara, CA, US), according to the manufacturer’s instructions.
Scan protocol
After 17 hours hybridization, slides were washed in staining dishes (Thermo Shandon, Waltham, MA, US) with Gene Expression Wash Buffer Kit (Agilent technologies, Santa Clara, CA, US) following the manufacturer’s instructions. Slides were scanned by Agilent Microarray Scanner (Agilent technologies, Santa Clara, CA, US) with default settings, Dye channel: Green, Scan resolution = 5 mm, PMT 100%, 16 bits.
Data processing
Data were extracted with 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). GSEA was performed following the developer’s protocol (http://www.broad.mit.edu/gsea/).
