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| Status |
Public on Apr 01, 2010 |
| Title |
Transcriptional profiles between mp mutant seedlings and transgenics carrying the dexamethasone-inducible GR-bdl protein |
| Organism |
Arabidopsis thaliana |
| Experiment type |
Expression profiling by array
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| Summary |
In order to identify targets of the transcription factor AUXIN RESPONSE FACTOR5 / MONOPTEROS (ARF5/MP), we compared transcriptomes of mp-B4149 mutant seedlings (9 day-old) and seedlings carrying the dexamethasone-inducible version of the MP inhibitor protein BODENLOS (GR-bdl). Without dexamethasone (DEX) treatment, this line is identical to the wild-type, while DEX treatment leads to strong inhibition of ARF-dependent transcription. To remove all endogenous MP-inhibiting Aux/IAA proteins, we treated mp or GR-bdl seedlings during 1 hour with auxin (50 micromolar Indole-3-Acetic Acid), either with or without a pretreatment with 10 micromolar DEX for 1 hour. Genes that are activated by MP are expected to br downregulated in mp seedlings and in the GR-bdl line afer DEX treatment. We used biological duplicates for each of the three treatments.
Keywords: Transcriptional profiling of transcription factor targets
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| Overall design |
Mp mutant and GR-bdl seeds were germinated on solid 1/2MS media, and transferred to liquid medium after 9 days of horizontal growth. Duplicate cultures were generated for mp mutants, and quadruplicates for the GR-bdl line. 10 micromolar DEX was added to both mp cultures and to two of the GR-bdl cultures, and seedlings were gently shaken during 1 hour. Then, 50 micromolar IAA was added to all 6 cultures and seedlings were incubated with gentle shaking for another hour. Total RNA was isolated from the seedlings using a QIAgen RNeasy kit, and reverse transcription, in-vitro transcription, cRNA labling and fragemntation was done according to established methods (Schmid et al., Nature Genetics [2005] 37, 501-506). Labeled cRNA was hybridized to Arabidopsis ATH1 arrays.
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| Contributor(s) |
Schlereth A, Juergens G, Weijers D |
| Citation(s) |
20220754 |
| Submission date |
Dec 09, 2008 |
| Last update date |
Jun 12, 2017 |
| Contact name |
Dolf Weijers |
| E-mail(s) |
dolf.weijers@wur.nl
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| Phone |
+31-317-482866
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| Fax |
+31-317-484801
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| URL |
http://www.bic.wur.nl
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| Organization name |
Wageningen University
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| Department |
Laboratory of Biochemistry
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| Street address |
Dreijenlaan 3
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| City |
Wageningen |
| ZIP/Postal code |
6703 HA |
| Country |
Netherlands |
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| Platforms (1) |
| GPL198 |
[ATH1-121501] Affymetrix Arabidopsis ATH1 Genome Array |
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| Samples (6)
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| Relations |
| BioProject |
PRJNA110351 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE13881_RAW.tar |
13.6 Mb |
(http)(custom) |
TAR (of CEL) |
| Processed data included within Sample table |
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