Project description:Single cell sequencing (DropSeq) from 7 day old Arabidopsis roots with β-estradiol-VND7, MYB83 and MYB46 induction lines and controls
Project description:Global transcriptome patterns were determined in XVE-14 and wild-type seedlings induced for 45 min b-estradiol in order to identify the genes early regulated by EBE transcription factor. We used microarrays to identify genes differentially expressed in EST-inducible EBE over-expression line #14 compared to wild-type plants, 45 min after 2µM EST induction. Three independent biological replications were performed. In order to identify potential direct/early target genes of EBE transcription factor, estradiol inducible overexpression system was used. Three week old Arabidopsis EBE-XVE (line 14) and WT seedlings were treated with ß-estradiol (2µM) for 45 min. RNA was isolated from shoot and subjected to hybridization on Affymetrix microarrays. Experiment was performed in 3 biological replications and genes differentially expressed between estradiol treated EBE-XVE and WT plants were identified as potential early targets of EBE.
Project description:Transcription profiling of seven-day-old seedlings of p35S:OBP4-GR treated with DEX or EtOH for 12 and 24 h. Purpose: find transcriptionally regulated genes downstream of the dof transcription factors OBP4 in the root tip of Arabidopsis thaliana Method: compare transcript levels after ectopic induction of OBP4 for 12 h and 24 h. Ectopic expressing was induced by introducing the coding sequence of OBP4 fused to the glucocorticoid receptor under the control of the Cauliflower Mosaic Virus p35S promoter (p35S:OBP4-GR) in wild type Arabidopsis Col-0 plants. Seven day old plants expressing the construct were treated with Dexamethason (DEX) or ethanol (EtOH) as a mock for 12 and 24 h. After root tips of 300 seedlings per rep were collected.
Project description:The experiment is designed to examine changes in gene expression upon ectopic expression of the germ line transcription factor DUO1 in Arabidopsis. Plants were transformed with the vector pMDC7DUO1, allowing induction of DUO1 by treatment with estradiol. T2 seedlings were selected on agar plates containing hygromycin for 12 days before seedlings were transferred to plates containing either 2 µM 17beta-estradiol (induced), or control plates containing DMSO (non-induced). Plants were grown for 6, 12 or 24 hours before harvesting. RNA was extracted using the RNAeasy kit from Qiagen. Each sample consists of 20 seedlings combined and all treatments have been conducted in triplicate.
Project description:Exploring miRNA-related antisense transcription in Arabidopsis through RNA transcript profiling of smRNA pathway-defective mutants on a custom high-resolution oligonucleotide array.
Project description:The experiment is designed to examine changes in gene expression upon ectopic expression of the germ line transcription factor DUO1 in Arabidopsis. Plants were transformed with the vector pMDC7DUO1, allowing induction of DUO1 by treatment with estradiol. T2 seedlings were selected on agar plates containing hygromycin for 12 days before seedlings were transferred to plates containing either 2 µM 17beta-estradiol (induced), or control plates containing DMSO (non-induced). Plants were grown for 6, 12 or 24 hours before harvesting. RNA was extracted using the RNAeasy kit from Qiagen. Each sample consists of 20 seedlings combined and all treatments have been conducted in triplicate. 18 samples were used in this experiment.
Project description:Arabidopsis Enhanced Disease Susceptibility 1 (EDS1) and its direct partner Phytoalexin Deficient 4 (PAD4) are required for both basal resistance against virulent pathogens and innate immune responses mediated by all tested TIR (Toll-Interleukin-1 Receptors) type nucleotide-binding/leucine-rich-repeat (NLR) receptors. Using transgenic Arabidopsis plants in Col-0 eds1-2 pad4-1 background conditionally expressing PAD4 and constitutively expressing EDS1 (ED-P4E1), an EDS1/PAD4 immune response can be triggered by estradiol treatment and can induce expression of defense-related genes and lead to enhanced basal resistance. In order to capture primary transcriptional changes in response to EDS1/PAD4, we performed RNA-seq gene expression analysis in ED-P4E1 plants after estradiol or mock treatment at three time points: 6, 12 and 24 h.
Project description:We defined the C/EBPa signature characterized by a set of genes which are upregulated upon C/EBPa activation. In order to identify the C/EBPa signature, we performed microarray gene expression analysis of K562 cells stably expressing p42-C/EBPa-ER after activating the C/EBPa construct to translocate to the nucleus for 6 hours with beta-estradiol. The gene expression profile was performed in K562 p42-C/EBPa-ER expressing cells treated with beta-estradiol (n=4) or EtOH vehicle control (n-4) for 6 hours. Induction of nuclear localization of C/EBPa-ER fusion protein was achieved by addition of 1 uM beta-estradiol as indicated into the culture medium.
Project description:Five members of the Arabidopsis thaliana NF-YA gene family are strongly induced by several stress conditions via transcriptional and miR169-related posttranscriptional mechanisms. These transcription factors participate in gene regulation via two different mechanisms, one depending on binding to the CCAAT-box in the promoter of regulated genes and the other, independent of the CCAAT-box, in which NF-YA prevents the interaction of the NF-YB/YC heterodimer with transcription factors. Three biological and two technical (in swap) replicates for each genotype were obtained for each treatment (DMSO (mock) and estradiol 24h after induction). Mock samples were pooled and used as a reference.
Project description:Transcriptional profiling of BJAB cells expressing miR-K12-6-5p. To identify host RNA targets of KSHV miRNAs, we took advantage of the observation that RNAs targeted by miRNAs often display small reductions in their steady-state levels, perhaps as a result of their impaired translation. Accordingly, we examined cellular transcript accumulation by array-based expression profiling under four sets of conditions in which KSHV miRNAs were expressed.