Treatment protocol
KM-H2-KLF4ER and L428-KLF4ER cells were incubated with 200 µM 4-OHT or with relevant amount of the vehicle (ethanol). The experiment was done twice. After 24 h of incubation total RNA was isolated with RNeasy mini kit (QIAGEN).
Growth protocol
KM-H2-FOXO1(3A)ER and L428-FOXO1(3A)ER cHL cells stably expressing FOXO1(3A)ER were grown in RPMI1640 medium with addition of 10% FCS, glutamine and antibiotics at 37C, 5% CO2
Extracted molecule
total RNA
Extraction protocol
Total RNA was isolated with RNeasy mini kit (QIAGEN) according to the manufacturers instruction.
Label
biotin
Label protocol
The total RNAs were amplified and labeled following the Whole Transcript (WT) Sense Target Labeling Assay (
http://www.affymetrix.com
).
Hybridization protocol
Labeled ssDNA was hybridized to Human Gene 1.0 ST Affymetrix GeneChip arrays (Affymetrix, Santa Clara, CA).
Scan protocol
The chips were scanned with a Affymetrix GeneChip Scanner 3000 according to the Affimetrix protocol.
Description
KM-H2-FOXO1(3A)ER cells incubated with vehicle for 24 hours
Data processing
The microarray data were processed using GeneSifter software (
http://www.geospiza.com
) using RMA normalization protocol.
