Project description:Mice were infected with Helicobacter pylori strain SS1 for 2, 7, 14 or 28 days or left uninfected. They were sacrificed at the appropriate times, the stomachs were removed and one half was plated for colony counts. The other half was embedded in OCT and cryosections were produced for 6-7 animals per timepoint. The sections were stained using the Histogene dye and roughly 500-1000 cells per gastric epithelial cell type were harvested by laser microdissection. RNA was prepared from these samples, subjected to two rounds of amplification, labelled and hybridized to 40.000 element murine cDNA arrays. The array names are composed as follows: "LCM" stands for laser capture microdissection; "mock" or "SS" stands for mock-infected or SS1-infected, respectively; "2,7,14 and 28" refers to the length of infection; "1-4" corresponds to the numbers given to every animal.The cellular origin of the RNA is represented by "mucus producing" or pit cell, "parietal" or acid-producing cell and "chief" or zymogenic cell. Between 5 and 7 samples were harvested per timepoint and cell type. Where the first labelling reaction and hybridization did not produce sufficiently good data (less than half of all spots made a regression correlation cutoff of >0.6), both were repeated (indicated by the addition of "relabeled" to the array name). The differential regulation of genes can be analyzed in both a time and cell type specific manner using publicly available software. Cell Type: "mucus producing" or pit cell (pit), acid-producing cell (parietal) and zymogenic cell (chief)
Project description:All arrays use the same reference, which is cDNA generated from RNA harvested from uninfected polarized T84 monolayers. Infections were done on two seperate days. Numbers following the name of each array designate the time point post H. pylori infection. Data for time course one is named such that only this timepoint is listed, while data for time course two (wildtype and mock) is named such that the time point is followed by the designation "-2". An all pairs experiment design type is where all labeled extracts are compared to every other labeled extract. Keywords: all_pairs
Project description:We obtained gene-expression profiles of microdissected renal-tubule cells from patients with proteinuric nephropathies. Based on the renal function during follow-up, the patients were divided in stable (n=14) and progressive (n=7) subjects (table 1). The cells of interest were laser-capture microdissected from frozen sections from archived kidney biopsy material. Initially all samples were processed as technical duplicates (2 x 21 arrays); due to a large number of signal-negative spots several arrays were excluded leaving 36 arrays for analysis. The samples P2, P6, P7, S10, S13 and S14 were analysed as individual arrays, all other samples were analysed after combination of duplicate arrays. Quality control: To test for reproducibility we calculated the intra-array variability of the duplicate arrays. Duplicate arrays were combined before statistical analysis where applicable. Patient and control characteristics can be found in the manuscript and on our website Frozen kidney biopsies were stained for alkaline phospatase, then the tubule cells were lasercapture microdissected using the PixCell II Laser Capture Microdissection System and CapSure; LCM Caps. Set of arrays that are part of repeated experiments Disease State: samples from patients with proteinuric nephropathy (stable or progressive)
Project description:The purpose of the experiment was to compare the transcriptional profile of lupus nephritis kidney tissue at a first flare and expression at a repeated lupus nephritis episode. All samples were laser microdissected into glomerular and tubular compartments and samples were ran in different cartridges.
Project description:Question Addressed: What is the host response (specifically apoptosis pathways) to Shigella infection and what is the contribution of MxiE to the host response? The arrays are apoptosis Exon hit arrays, thus splice variation can be determined. Array data from HeLa cells that remained Uninfected or were infected with wildtype S. flexneri serotype 2a strain 2457T or an isogenic mixE mutant. These groupings are further divided such that Uninfected or Infected cells remained untreated or were treated with the apopotosis inducer staurosporine (STS). The design of the experiment was a time course and samples were harvested at the indicated time points (hr). All hybridizations were conducted against a common reference sample that was made from pooled samples of normal uninfected healthy HeLa cells. Infection: HeLa cells were infected with wt Shigella (wt), mxiE mutant (mixE) or were left uninfected (uninfected) Compound Based Treatment: Cultures were treated with Staurosporin (STS) or were left untreated (untreated) Infection time: Cells were harvested at the indicated time after treatment
Project description:Mice were infected with Helicobacter pylori strain SS1 for 2, 7, 14 or 28 days or left uninfected. They were sacrificed at the appropriate times, the stomachs were removed and one half was plated for colony counts. The other half was embedded in OCT and cryosections were produced for 6-7 animals per timepoint. The sections were stained using the Histogene dye and roughly 500-1000 cells per gastric epithelial cell type were harvested by laser microdissection. RNA was prepared from these samples, subjected to two rounds of amplification, labelled and hybridized to 40.000 element murine cDNA arrays. The array names are composed as follows: "LCM" stands for laser capture microdissection; "mock" or "SS" stands for mock-infected or SS1-infected, respectively; "2,7,14 and 28" refers to the length of infection; "1-4" corresponds to the numbers given to every animal.The cellular origin of the RNA is represented by "mucus producing" or pit cell, "parietal" or acid-producing cell and "chief" or zymogenic cell. Between 5 and 7 samples were harvested per timepoint and cell type. Where the first labelling reaction and hybridization did not produce sufficiently good data (less than half of all spots made a regression correlation cutoff of >0.6), both were repeated (indicated by the addition of "relabeled" to the array name). The differential regulation of genes can be analyzed in both a time and cell type specific manner using publicly available software. Cell Type: "mucus producing" or pit cell (pit), acid-producing cell (parietal) and zymogenic cell (chief) time_series_design
Project description:Adult male Sprague Dawley rats (5 month-old at the beginning of the study) were subcutaneously exposed to 0.5 mg.kg/d rotenone or the corresponding solvent (PEG/DMSO, 1/1) for 28 days. At the end of the experiment, animals were sacrificed by decapitation and brains were rapidly taken out within 30-40 s. Total RNA was prepared from laser-capture microdissected rat substantia nigra pars compacta (Snpc). Changes in transcript abundance within SNpc were analyzed on samples from solvent and rotenone-exposed groups (n = 3 per group) using CodeLink™ Rat Whole Genome bioarrays containing 34,000 transcripts and EST probes (GE Healthcare, Amersham, Saclay, France).
Project description:We sequenced cDNA synthesized from mRNA from 3 time points of a time course of Arabidopsis thaliana shoot apical meristems. Plants were grown two weeks in short days (time point +0LD) and then exposed to long days for 1 day (time point +1LD) and 3 days (time point +3LD). Meristem cells were collected using laser capture microdissection, to generate a meristem-specific time course of gene expression in the early stages of floral induction. The experiment was done in three biological replicates (called A, B, and C). Some repetitions of sequencing on a few samples (technical replicates) were also done
Project description:The microarrays for the three time course experiments described in the publication titled "Gene expression profiling of Helicobacter pylori reveals a growth phase dependent switch in virulence gene expression". TC1 refers to the first time course investigating gene expression changes over the time course of growth while TC2 refers to the second time course. TC Motility_broth refers to the third time course described in the manuscript used to investigate the concurrent changes in motility and the expression of the genes in the flagellar regulon. Detailed description of the growth of H. pylori for these 3 time courses can be found in the Materials and Methods section of the manuscript along with the exact filtering criteria used to download the data from these arrays. Note that in all three time courses the data for each array were transformed after download such that the abundance of each gene's transcript represented by a given spot was relative to the level of that transcript at the 6 h time point. Also duplicate spots for each ORF on the microarray were averaged for analysis after transformation. Groups of assays that are related as part of a time series. Keywords: time_series_design