Transcription profiling of wild type and rcd3-1 mutant Arabidospsi plants
Ontology highlight
ABSTRACT:
INSTRUMENT(S): other
ORGANISM(S): Arabidopsis thaliana
SUBMITTER: Mikael Brosche
PROVIDER: E-MEXP-1388 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:23 day old Arabidopsis plants, wildtype Col-0 and mutant rcd1, were compared to determine which genes are over- or under expressed in the mutant. Six biological repeats were pooled in pairs of two for labellings. All labellings/hybridizations were done with a dye swap technical repeat (e.g. in total six labellings were used). The Turku Arabidopsis 8K arrays were used for hybridizations. For the hybridizations the samples were split in two halves with each half hybridized to Turku Arabidopsis 8K slide 1 and Turku Arabidopsis 8K slide 2 (e.g. in total 12 hybridizations were performed).
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2008-12-12 | E-MEXP-1949 | biostudies-arrayexpress
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2008-01-11 | E-GEOD-10145 | biostudies-arrayexpress
Project description:Mouse macrophages J774A.1 were pre-treated with the anti-inflammatory lipid eicosapenaenoic acid (EPA) or the pro-inflammatory lipid ceramide (Cer) for 3h. Macrophages were then infected with Mycobacterium smegmatis for 1h, and total RNA was collected. In a parallel experiment, infected macrophages were infected for 1h, 4h and 24h. At these time points, macrophages were lysed, bacteria was collected and quantified by the Colony Forming Units (CFU) Assay. This provided the kinetics of the killing of Mycobacteria inside mouse macrophages. CFU experiments revealed that cells pre-treated with EPA showed an increased number of bacteria inside macrophages, in contrast to cells pre-treated with Cer. To dissect the molecular mechanisms involved in the survival and killing of mycobacteria infected macrophages, mediated by lipids, gene expression studies were performed. Cultures of Mycobacterium smegmatis mc2155 harbouring a p19-(long lived) EGFP plasmid were grown to exponential growth phase. Bacteria were pelleted, washed in PBS and resuspended in medium DMEM with a multiplicity of infection (MOI) of 10 (10 bacteria per macrophage). Clumps of bacteria were removed by ultrasonic treatment of bacterial suspensions in an ultrasonic water bath for 15 minutes, followed by a low speed centrifugation for 2 minutes. Mouse macrophages J774A.1 were pre-.treated with the anti-inflammatory lipid eicosapenaenoic acid (EPA) or the pro-inflammatory lipid ceramide (Cer), 3h before infection. Mouse macrophages J774A.1 were infected with Mycobacterium smegmatis mc2155 for 1h, at 37M-BM-:C and 5% CO2. After 1h of infection, cells were washed with PBS. After washing, 1 ml Trizol was added per well, to collect total RNA. The experimental condition were: samples 1.1, 1.2, 1.3: Untreated macrophages; samples 2.1, 2.2, 2.3: Mock treated macrophages (ethanol 1ul/ml); samples 3.1, 3.2, 3.3: EPA (15uM) treated macrophages (ethanol 1ul/ml); samples 4.1, 4.2, 4.3: Cer (5ug/ml) treated macrophages (ethanol 1ul/ml); samples 5.1, 5.2, 5.3: Untreated macrophages, infected with Mycobacterium smegmatis mc2155 for 1h; samples 6.1, 6.2, 6.3: Mock treated macrophages (ethanol 1ul/ml) infected with Mycobacterium smegmatis mc2155 for 1h; samples 7.1, 7.2, 7.3: EPA (15uM) treated macrophages (ethanol 1ul/ml) infected with Mycobacterium smegmatis mc2155 for 1h; samples 8.1, 8.2, 8.3: Cer (5ug/ml) treated macrophages (ethanol 1ul/ml) infected with Mycobacterium smegmatis mc2155 for 1h; In a parallel experiment, infected macrophages remained with the bacteria for 1h, 4h and 24h after infection. After these time points, the macrophages were lysed, the bacteria was collected and quantified by the Colony Forming Units (CFU) Assay. This provided the kinetics of the killing of non-pathogenic intracellular bacteria inside mouse macrophages. For the CFU assay, after 1h of infection, cells were washed with PBS and Gentamicin (10ug/ml) in DMEM to kill the extracellular bacteria. At discrete time points, cells were washed with PBS and lysed with sterilized water. Quantitative cultures of bacteria were performed in a 10-fold serial dilutions, inoculated on 7H10 agar plates. 5ul were plated in triplicate and the number of colonies were counted after 48h.
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2007-08-01 | E-MEXP-1053 | biostudies-arrayexpress
Project description:22 day old Arabidopsis Col-0 plants were sprayed with 0.5 mM sodium nitroprusside (SNP), corresponding controls were sprayed with water, and leaves harvested 3 hours after spraying and frozen in liquid nitrogen.
2007-09-28 | E-MEXP-1268 | biostudies-arrayexpress
Project description:We are studying the tree P. euphratica growing in its natural habitat in the Negev desert in Israel. We have used leaf RNA samples from trees growing from four different areas in the desert valley Ein Avdat with contrasting growth conditions, with the primary factor being how much water the trees have access to. Area A trees grow close to a stream. Area B trees are further away from the stream. Area C trees are growing on a slope with no water. Parking place trees grow at a parking place (1km from Area A, B and C) where there is a water irrigation system, and these trees are regularly watered once a week. The control sample in each hybridization is always RNA isolated from a pool of 5 trees from the parking place. The hybridizations are comparing the following: Area A -- parking place Area B -- parking place Area C -- parking place Each hybridization is done with a dye swap and three different biological repeats for a total of 18 hybridizations.
2004-11-30 | E-MEXP-182 | biostudies-arrayexpress
Project description:Rooted plantlets of Populus euphratica were transferred to aerated hydroculture with Long Ashton nutrient solution, grown for 3.5 months at 22°C, 150 µmol m-2 s-1 photosynthetically active radiation with a photoperiod of 16h light. After reaching an average height of 0.83 m, the media was supplemented with 150 mM NaCl (Salt-treated) or maintained under control conditions (control). During the experimental phase, light was continuously supplied to avoid interference of light/dark transitions. <br>Roots and leaves of six plants were harvested separately after 3h, 6h, 12h, and 24h of salt exposure and of controls at the corresponding time points (= 48 plants per experiment). The material of each harvest of 6 plants was pooled, yielding one leaf and one root sample of controls and one leaf and one root sample of salt-treated plants, respectively (=16 samples per experiment).<br>The experiment was performed three times yielding three biological replications (=48 samples in total).<br>After extraction of RNA and cDNA synthesis, each extract was labelled with Cy3 and Cy5, respectively (yielding 96 labelled extracts). Extracts from e.g. 3h-salt-treated leaf exp. 1 (cy3) was hybridized with 3h-control leaf exp.1 (cy5) with microarrays (UHPB-P.euphratica-10K-5; A-MEXP-93)containing unique 7,662 ESTs of P. euphratica from stress-induced cDNA libraries (Brosche et al., Genome Biology 2005, 6:R101). Dye swap was perfomed, thus usually yielding 6 arrays per timepoint and plant tissue (3 biological, 3 technical replicates). In some cases hydridizations were not successfull. Therefore, for roots harvested at timepoints 12h and 24 h only 4 arrays per timepoint were used. For roots harvested after 3h an additional mixed sample of all 3 biological replicates was performed. One leaf array at 6 h was not sucessfull. Therefore, residual labelled extract from salt treated plants of exp1 was hydridized with controls at 6h from exp.2. <br>
2009-08-05 | E-MEXP-2095 | biostudies-arrayexpress
Project description:Tissues of Arabidopsis plants overexpressing artificial microRNAs were compared to wild_type and respective target gene mutants (duplicate arrays)
2006-04-01 | E-TABM-63 | biostudies-arrayexpress
Project description:Plantlets of P. euphratica (clone B2) were obtained by in vitro culture. After ex vitro acclimation, they were transferred and acclimated to greenhouse conditions and transplanted into 7.5L-pot filled with peat-sand mix (50/50 V/V) and were pruned (following some leaf shedding due to transplantation shock). In the greenhouse, climate conditions depended on outside weather but temperature was regulated in order to remain between 15 and 30°C. A moderate, increasing drought stress was applied and controlled for 6 weeks. Soil volumetric water content was measured one to two times a day depending on the intensity of the stress (TDR and weighting). Plantlets were progressively brought to 5 stress levels [soil volumetric water content]: 10% (harvest 1, 11 days from start of experiment), 7.5% (harvest 2, 16 days from start of experiment), 5% (harvest 3, 22 days from start of experiment), 4% (harvest 4, 26-32 days from start of experiment) and back to fully available water (harvest 5, 10 days after fully watering of the trees; 36 days from start of experiment). Leaves and roots (including control and stressed trees) were harvested for each stress level. At harvest 1-3, samples from five trees were harvested. At harvest 4, samples from two trees were harvested at each of day 26, 29 and 32. At harvest 5, samples were harvested from two control trees and four re-watered trees. <br><br> For the array experiment: leaf or root samples were pooled before RNA isolation to give three repeats by dividing the five samples 2+2+1. For harvest 4; samples were pooled from each of the two trees sampled at day 26, 29 and 32 to give three biological repeats. In harvest 5 samples were pooled 2+2 to give two biological repeats. Each array experiment was done with a dye swap to give 28 hybridizations for the leaf and 28 hybridizations for the root experiment.
2005-03-01 | E-MEXP-276 | biostudies-arrayexpress