cell line: HepG2 cell type: Cell culture time of exposure: 24 hours agent: 2,4-D dose (um): 0.03
Treatment protocol
Cells were exposed in 96-well plates 24hrs after plating to allow attachment. 2,4-dichlorophenoxyacetic acid, fenbuconazole, and imazalil were purchased from Sigma-Aldrich (St. Louis, MO). Stock solutions of 2,4-D, fenbuconazole, and imazilil were diluted in DMSO and added to culture media to a final DMSO concentration of 0.2%. The cells were exposed for 24hrs to the DMSO vehicle (0.2%) or one of the following chemical concentrations: 0.001, 0.003, 0.01, 0.03, 0.1, 0.3, 1, 3, and 10uM.
Growth protocol
HepG2 cells were purchased from ATCC (Manassas, VA). HepG2 cells were cultured in DMEM (cellgro) containing 10% FBS, 1X pen/strep, and 1X NEAA and plated at 18,000 cells/well in 96 well plates.
Extracted molecule
total RNA
Extraction protocol
Trizol® (ThermalFisher) organic extraction with Phase Lock Gel™ -Heavy- tubes (5Prime). Aqueous phase placed in RB(QiaCube sample) tubes and loaded into QiaCube, the RNeasy Mini Protocol used.
Label
biotin
Label protocol
Affymetrix, GeneChip® High Throughput 3' IVT PLUS Reagent Kit
Hybridization protocol
Affymetrix, GeneChip® High Throughput 3' IVT PLUS Reagent Kit, page40-42 and GeneTitan® Instrument User Guide for Expression Array Plates
Scan protocol
Arrays were scanned on an Affymetrix GeneTitan array scanner to generate CEL files with corresponding intensity values.
Description
Gene Expression data from HepG2 cell culture BW_003_24h_259_P1_2-4-D-HepG2
Data processing
Affymetrix CEL files were normalized by RMA using Partek Genomic's Suite 6.6 (each group normalized independently)
