cell line: Hep3b cell type: Liver cancer cells agent: CU27
Treatment protocol
The cells were washed with PBS after removed medium, and the TriZol was added into the cell monolayer, gently mixed with the tips, and immmediately freezed with liquid N2, followed by stored in the -80 oC freezer.
Growth protocol
The cells were maintained in DMEM (Sigma) or MEM (Sigma) containing 10% FBS (ExCell Biology, Shanghai, China) at 37°C with 5% CO2.
Extracted molecule
total RNA
Extraction protocol
Total cellular RNA was isolated from the cells using an RNeasy Mini Kit (Qiagen, Hilden, Germany) in accordance with the manufacturer’s protocol.
Label
Biotin
Label protocol
Total RNA were amplified, labeled and purified by using GeneChip 3’IVT Express Kit (Cat#901229, Affymetrix, Santa Clara, CA, US) followed the manufacturer’s instructions to obtain biotin labeled cRNA.
Hybridization protocol
Array hybridization and wash was performed using GeneChip® Hybridization, Wash and Stain Kit (Cat#900720, Affymetrix, Santa Clara, CA, US)in Hybridization Oven 645 (Cat#00-0331-220V, Affymetrix, Santa Clara, CA, US) and Fluidics Station 450 (Cat#00-0079, Affymetrix, Santa Clara, CA, US) followed the manufacturer’s instructions
Scan protocol
Slides were scanned by GeneChip® Scanner 3000 (Cat#00-00212, Affymetrix, Santa Clara, CA, US) and Command Console Software 3.1 (Affymetrix, Santa Clara, CA, US) with default settings.
Description
Gene expression data from hep3b cells treated with 10 μM CU27 (48 hours). hep3b-CU27-2
Data processing
Raw data were normalized by MAS 5.0 algorithm, Gene Spring Software 11.0 (Agilent technologies, Santa Clara, CA, US).
