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RNA-seq of Arabidopsis 14 day-old seedlings expressing either mCherry-RAX1-GR or mCherry-GR with 4h mock vs. dexamethasone treatment

ABSTRACT: RNA-seq of Arabidopsis 14 day-old seedlings expressing either mCherry-RAX1-GR or mCherry-GR with 4h mock vs. dexamethasone treatment

PROVIDER: PRJEB27846 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR2706900_1.fastq.gz	Fastqsanger.gz
	ERR2706900_2.fastq.gz	Fastqsanger.gz
	ERR2706901_1.fastq.gz	Fastqsanger.gz
	ERR2706901_2.fastq.gz	Fastqsanger.gz
	ERR2706902_1.fastq.gz	Fastqsanger.gz

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RNA-seq of Arabidopsis 14 day-old seedlings expressing either mCherry-RAX1-GR or mCherry-GR with 4h mock vs. dexamethasone treatment

Project description:In order to identify relevant target genes of RAX1 (AT5G23000) involved in meristem initiation in Arabidopsis we generated dexamethasone inducible lines expressing either a mCherry-RAX1-GR or a mCherry-GR (mock construct) fusion protein. We analysed differential gene regulation after 4 hours of dexamethasone or mock treatment in 14 day-old seedlings. Paired-end sequencing was performed using 3 biological replicate for each genotype and differentially expressed genes were identified for the interaction of genotype and treatment.

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Transcriptional profiles between mp mutant seedlings and transgenics carrying the dexamethasone-inducible GR-bdl protein

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Transcriptional profiles between mp mutant seedlings and transgenics carrying the dexamethasone-inducible GR-bdl protein

Project description: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 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.

2010-04-12 | E-GEOD-13881 | biostudies-arrayexpress

Expression data from Arabidopsis GR-REVOLUTA and KANADI1-GR transgenic seedlings

Project description:The REVOLUTA and KANADI1 transcription factors act in opposite directions to control polarity in Arabidopsis. By analyzing mRNA profiles following activation of REVOLUTA or KANADI1 proteins we identified genes regulated by both transcription factors, focusing on those regulated in opposite directions. To coordinate transcriptional responses for microarray analysis, we constructed transgenic seedlings with the REV or KAN1 transcription factor fused to a glucocorticoid receptor (GR) domain. The GR domain tethers the transcription factor in the cytoplasm, and treatment with dexamethasone (DEX) allows movement into the nucleus. Transgenic seedlings and wild-type controls were grown simultaneously in liquid culture, treated with dexamethasone for 0min, 30min, 60min, or 120min, and total RNA was extracted and hybridized to Affymetrix arrays. Treatment: Dexamethasone Biological replicates

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Response to1 µM dexamethasone in MEF cells expressing GR dimer deficient mutant receptors

Project description:The glucocorticoid (GC) receptor (GR) is essential in development and inflammation. Healthy GRdim/dim mice with reduced dimerization propensity due to a point mutation (A465T) at the dimer interface of the GR DNA binding domain (DBD) (here GRD/D) have helped to define GR monomer and dimer functions of GR. Since GRD/D retains residual dimerization capacity, we generated the dimer-nullifying double mutant GRD+L/D+L mice, featuring an additional mutation (I634A) in the ligand binding domain (LBD) of GR. These mice are perinatally lethal, as are GRL/L mice, displaying improper lung and skin formation. We used embryonic fibroblasts, high and low doses of dexamethasone (Dex), nuclear translocation, RNAseq, dimerization assays and ligand binding assays (and Kd values), we suggest that the lethal phenotype is due to insufficient ligand binding. The data suggest cross talk between GR dimerization potential and ligand affinity. We conclude that a mutation as subtle as this I634A, at a position not directly involved in ligand interactions senso stricto, can thus still influence ligand binding and have a lethal outcome.

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E-MTAB-7050

Project description:RNA-seq of Arabidopsis 14 day-old seedlings expressing either mCherry-RAX1-GR or mCherry-GR with 4h mock vs. dexamethasone treatment

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Whole-genome GR binding sites and histone acetylation status in pediatric acute lymphoblastic leukemia patient-derived xenografts following dexamethasone treatment in vivo

Project description:Glucocorticoids are critical components of combination chemotherapy regimens in pediatric acute lymphoblastic leukemia (ALL). However, the signaling pathways regulating apoptosis in glucocorticoid-treated lymphoid cells remain unclear. In this study, pediatric ALL patient-derived xenograft inherently sensitive to glucocorticoids were exposed to dexamethasone in vivo. Whole-genome GR binding sites and histone acetylation status were detected using chromatin immunoprecipitation sequencing analyses. This provided a global understanding of dexamethasone-induced DNA modulations in ALL cells in vivo, which is likely to be important in the understanding of mechanisms of glucocorticoid response in lymphoid malignancies. One xenograft (ALL-54) was derived from a patient of dexamethasone-good responder. The xenograft was innoculated into 2 NOD/SCID mice, and treated with dexamethasone (15 mg/kg) or vehicle control. Binding of glucocorticoid receptor (GR), histone acetylation and IgG control in spleen-harvest xenograft samples were detected using ChIP-seq.

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Dexamethasone stimulated gene expression in peripheral blood

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