KSHV microRNA transfection and human mRNA expression changes
ABSTRACT: Transcriptional profiling of HUVECs transfected with KSHV miRNA mimics (vMix) or a negative control mimic (neg). To identify host RNA targets of KSHV miRNAs, we took advantage of the observation that RNAs targeted by miRNAs can display small reductions in their steady-state levels. Cells transfected with negative control miRNA compared to cells transfected with KSHV miRNAs. Experimental treatment versus control treatment
Project description:Transcriptional profiling of BJAB cells expressing miR-K5 and BCBL cells treated with miR-K5 inhibitor. 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 or inhibited. Keywords: Experimental treatment versus control treatment Cells transfected with negative control miRNA compared to miR-K5 or negative miRNA inhibitor compared to miR-K5 inhibitor or stable cells transduced with empty vector compared to stable cells transduced to express miR cluster K1,2,3,4,5.
Project description:The experiment followed transcriptional changes during potato tuber induction from a stolon tip to a tuber. Samples were taken at stage 1, stage 3, stage 4 and stage 5 according to Kloosterman et al., 2005
Project description:To compare the expression profile of differentiated mouse bone marrow macrophages (BMM) in response to IL-4, we have employed whole genome microarray expression profiling. For this purpose, bone marrow cells were isolated from 8 to 12 weeks old C57BL6/J and Balb/cAnNCrl mice and cultured in the presence of the macrophage colony-stimulating factor (M-CSF). After seven days of culture, IL-4 was added for 4 and 18 hours. Keywords: Mice strain comparison; Gene expression profiling IL-4 induced gene expression was investigated in mouse bone marrow macrophages (BMM) of C57BL6/J and Balb/cAnNCrl mice. Differentiated BMM were incubated with mouse recombinant IL-4 for 4 or 18 hours or without for 18 hours. Two independent experiments were performed at each time (mock, 4 and 18 hours) using different mice littermates for each experiment.
Project description:Microarray analysis was performed on in vivo-derived RNA from the well characterized GAS strain 5448 and its derivative 5448R-, containing the mutated ropB allele from 5628. We followed a cyclic design that allowed us to compare every condition to each other at least twice, and guaranteed dye swapping to eliminate effects of non-specific binding. This study consists of 4 replicates of 5448 and 3 replicates of 5448R-.
Project description:Background Small colony variants (SCVs) are slow-growing bacteria, which often show increased resistance to antibiotics and cause latent or recurrent infections. It is therefore important to understand the mechanisms at the basis of this phenotypic switch. Methodology/Principal findings One SCV (termed PAO-SCV) was isolated, showing high resistance to gentamicin and to the cephalosporine cefotaxime. PAO-SCV was prone to reversion as evidenced by emergence of large colonies with a frequency of 10-5 on media without antibiotics while it was stably maintained in presence of gentamicin. PAO-SCV showed a delayed growth, defective motility, and strongly reduced levels of the quorum sensing Pseudomonas quinolone signal (PQS). Whole genome expression analysis further suggested a multi-layered antibiotic resistance mechanism, including simultaneous over-expression of two drug efflux pumps (MexAB-OprM, MexXY-OprM), the LPS modification operon arnBCADTEF, and the PhoP-PhoQ two-component system. Conversely, the genes for the synthesis of PQS were strongly down-regulated in PAOSCV. Finally, genomic analysis revealed the presence of mutations in phoP and phoQ genes as well as in the mexZ gene encoding a repressor of the mexXY and mexABoprM genes. Only one mutation occurred only in REV, at nucleotide 1020 of the tufA gene, a paralog of tufB, both encoding the elongation factor Tu, causing a change of the rarely used aspartic acid codon GAU to the more common GAC, possibly causing an increase of tufA mRNA translation. High expression of phoP and phoQ was confirmed for the SCV variant while the revertant showed expression levels reduced to wild-type levels. Conclusions By combining data coming from phenotypic, gene expression and proteome analysis, we could demonstrate that resistance to aminoglycosides in one SCV mutant is multifactorial including overexpression of efflux mechanisms, LPS modification and is accompanied by a drastic down-regulation of the Pseudomonas quinolone signal quorum sensing system. We used microarrays to study changes in gene expression during early and late stationary phase of SCV and WT strains. SCV and WT cultures were grown in triplicate until early (24h) and late (48h) stationary phase. RNA was extracted, labelled and hybridised on Affymetrix P. aeruginosa expression microarrays.
Project description:This SuperSeries is composed of the following subset Series: GSE16353: The profile of cellular and KSHV microRNAs in AIDS_KS biopsies (and normal skin control biopsies) GSE16354: Infection of Lymphatic and Blood Vessel Endothelial Cells (LEC and BEC) with KSHV GSE16355: Lymphatic endothelial cells (LEC) transfected with the KSHV microRNA cluster GSE16356: Lymphatic endothelial cells (LEC) treated with a MAF-targeted siRNA Refer to individual Series