Affymetrix GeneChip Mouse Genome 430A 2.0 [Mouse430A_2]

PU.1 Activation Relieves GATA-1-Mediated Repression of Cebpa and Cbfb during  Leukemia Differentiation

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17b-Estradiol added to MEL cells expressing Gata1-ER or PU.1-ER transgenes to stimulate either erythropoietic Gata-1 dependent or myeloid PU.1 dependent gene espression in different time points

GENOTYPE

TIME AFTER INDUCTION

Transcription profiling of mouse erythroleukemia cells following activation of Gata1-ER or PU.1-ER transgenes

17b-Estradiol added to MEL cells expressing Gata1-ER or PU.1-ER transgenes to stimulate either erythropoietic Gata-1 dependent or myeloid PU.1 dependent gene espression in different time points


P-AFFY-6

Title: Affymetrix CEL analysis. Description:

P-MTAB-4663

Mouse erythroleukemia cells (MEL) and HeLa cells were cultured in DMEM medium supplemented with 10% fetal bovine serum and antibiotics as described previously (Rekhtman 2003, Stopka 2005 and Rao 1997).

P-MTAB-4664

The PUER and GER conditional activation in MEL cells containing these transgenes (MELPUER and MELGER) was induced by 10-7 M 17b-Estradiol for indicated timepoints (0, 2, 4, 8, 12, 16, 20, 24hours).

P-MTAB-4665

Total cellular RNA was purified by a modified TRIzol reagent (Invitrogen) involving an enhanced precipitation by adding 1 volume of isopropanol to the extracted aqueous phase, precipitating at -20OC overnight, and centrifuging the RNA for 30 minutes at 14,000 rcf at 4OC. The concentration, purity and integrity of total RNA were determined by NanoDrop ND-1000 and Agilent 2100 Bioanalyzer.

P-MTAB-4666

one-cycle labeling protocol recommended by Affymetrix.

My scanning protocol

The microarrays were scanned using the GenePix 4200A scanner. GenePix Pro 5.1 software was used to determine the average signal intensity and the local background for each spot.

Transcription profiling of mouse erythroleukemia cells following activation of Gata1-ER or PU.1-ER transgenes

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