Project description:Comprehensive gene expression profiles of seven NK cell lines and eleven clinical samples of NK cell neoplasm were analyzed to detect tumor-specific genes. The gene expression profiles of NK cells from normal donor were analyzed as control samples.
Project description:Generation of monocyte-derived dendritic cells (DC). Immature DC were prepared from peripheral blood mononuclear cells (PBMCs) isolated from whole blood of healthy donors. A minimum of 3 individual donor blood was used for each experiment. PBMCs were incubated with anti-CD14 microbeads (Miltenyi Biotec, Bergish Gladbach, Germany) as specified in the manufacturer's instructions. The isolated monocytes were resuspended at 10^6 cells/ml in medium supplemented with 10% human AB serum, 100 ng/ml GM-CSF (Leucomax; Novartis, Camberley, UK) and 50 ng/ml IL-4 (Peprotech EC Ltd, London, UK), cultured in tissue culture flasks. On day 7 the cells were defined as immature DC with low/intermediate HLA-DR expression. Microarray analysis of the effects of Paclitaxel and LPS on DC. Total RNA was extracted from day 7 DC which had been treated for 2 h with 1 ug/ml LPS or 100 uM paclitaxel for 2 h then washed and returned to culture for 24 h. This was used as a template to generate Cy3-labelled cRNA, using the Low RNA Input Linear Amplification Kit (Agilent). This was used as a probe on the Whole Human Genome Microarray (4x44K) slide (Agilent). Slides were scanned using the Agilent scanner and data extracted using Feature Extraction Software 9.5.3 (Agilent). Subsequent data analysis was performed using GeneSpring GX software.
Project description:BSA pooling experiment for methionine content in a segrating diploid potato population (CxE). RNA of constrasting individuals for methionine content are pooled together based on their tuber methionine content and marker association with either or both of the identified QTLs for methionine content
Project description:Members of the SR protein family of RNA binding proteins play numerous roles in gene expression, including the regulation of pre-mRNA splicing, mRNA export, and translation. How SR proteins coordinate gene expression programs is unknown, and comprehensive knowledge of endogenous mRNA targets is lacking. We established physiological expression of GFP-tagged SR protein, allowing the immunopurification and systematic identification of mRNAs associated with SRp20 and SRp75. This genome-wide analysis showed that SRp20 and SRp75 are components of distinct, mostly non-overlapping mRNPs in undifferentiated and neural cells.
Project description:Members of the SR protein family of RNA binding proteins play numerous roles in gene expression, including the regulation of pre-mRNA splicing, mRNA export, and translation. How SR proteins coordinate gene expression programs has been largely unexplored, and comprehensive knowledge of endogenous mRNA targets is lacking. Gene expression changes upon SRp20 or SRp75 RNAi in P19 parental, SRp75-BAC, and SRp20-BAC cells were measured with mouse whole genome microarrays. The knockdown of either SRp20 or SRp75 led to up- or downregulation of specific transcripts. Phenotypic rescue in the SRp75-BAC, and SRp20-BAC cells demonstrated functionality of the GFP-tagged SR proteins, and showed the specificity of the knockdowns.
Project description:Transcriptional profiling of roots and hypocotyls of soybean comparing control untreated 2-d-old seedlings with flooding treated 2-d-old seedlings.
Project description:Background<br>Primitive brain tumors are the first cause of cancer-related death in children. Tumor cells with stem-like properties (TSCs), thought to account for tumorigenesis and therapeutic resistance, have been isolated from high-grade gliomas in adults. Whether TSCs are a common component of pediatric brain tumors and are of clinical relevance remains to be determined. <br>Methodology/Principal findings<br>Tumor cells with self-renewal properties were isolated with cell biology techniques from a majority of 55 pediatric brain tumors samples, regardless of their histopathologies and grades of malignancy (57% of embryonal tumors, 57% of low-grade gliomas and neuro-glial tumors, 70% of ependymomas, 91% of high-grade gliomas). The vast majority (10/12) of high-grade glioma-derived oncospheres sustained long-term self-renewal numbers akin to neural stem cells (>7 self-renewals), whereas cells with limited renewing abilities akin to neural progenitors dominated in all other tumor types. Regardless of tumor entities, the young age group was associated with self-renewal properties akin to neural stem cells (P=0.05, chi-square test). TSCs shared a complex molecular profile combining embryonic stem cell markers with elements controlling neural stem cell properties and epithelio-mesenchymal transitions. They were radio- and chemoresistant and formed aggressive tumors after intracerebral grafting. Survival analysis of the cohort showed an association between isolation of TSCs with long-term self-renewal abilities and a patients higher mortality rate (P = 0.022, log-rank test). Patients bearing cells with limited self-renewal properties constituted an intermediate group of survival but which did not reach statistical significance. <br>Conclusions/Significance<br>In brain tumors affecting adult patients, TSC have been isolated only from high-grade gliomas. In contrast, our data show that tumor cells with stem cell-like or progenitor-like properties can be isolated from a wide range of histological sub-types and grades of pediatric brain tumors. They suggest that cellular mechanisms fueling tumor development differ between adult and pediatric brain tumors.<br>
Project description:Transcriptional profiling of e8.5 mouse embryos comparing wt with Srd5a3Gt(betaGeo)703Lex/Gt(betaGeo)703Lex. Goal was to determine the developmental pathway disrupted in the mutant.<br>