Project description:[Hela cells]: We performed cdr2 knockdown with a pool of 4 cdr2-specific siRNAs to test whether cdr2 may regulate c-myc target genes as cells passage through mitosis. [Rat1a wild type and myc null cells]: We performed cdr2 knockdown using a pool of 4 cdr2-specific siRNAs to test whether cdr2 may regulate c-myc target genes as cells passage through mitosis.
Project description:[Hela cells]: We performed cdr2 knockdown with a pool of 4 cdr2-specific siRNAs to test whether cdr2 may regulate c-myc target genes as cells passage through mitosis. [Rat1a wild type and myc null cells]: We performed cdr2 knockdown using a pool of 4 cdr2-specific siRNAs to test whether cdr2 may regulate c-myc target genes as cells passage through mitosis. [HeLa cells]: Cells were transfected with control or cdr2 siRNAs and then cells were synchronized in mitosis using a sequential thymidine/nocodazole block. Cells were subsequently released from nocodazole block and after 3 hours RNA was harvested for microarray analysis [Rat-1 wild type (TGR) and c-myc null (15.19) cells]: Cells were transfected with control or cdr2 siRNAs and then cells were synchronized in mitosis using a sequential thymidine/nocodazole block. Cells were then subsequently released from nocodazole block and after 3 hours RNA was harvested for microarray analysis
Project description:T cell clonal expansion and differentiation are critically dependent on the transcription factor c-Myc (Myc). Herein we use quantitative mass-spectrometry to reveal how Myc controls antigen receptor driven cell growth and proteome restructuring in CD4+ and CD8+ T cells. Quantitative proteomics was performed on naive wild-type (WT) and 24 hr T cell receptor (TCR) activated Myc WT and Myc-deficient T cells. Analysis of copy numbers per cell of >7000 proteins provides new understanding of the selective role of Myc in controlling the protein machinery that shapes T cell fate. The data identify both Myc dependent and independent metabolic processes in immune activated T cells. We uncover that a primary function of Myc is to induce amino acid transporter expression. Quantitative proteomics of 24 hr TCR activated Slc7a5 WT and Slc7a5-deficient CD4 T cells reveals that loss of a single Myc-controlled amino transporter, Slc7a5, can effectively phenocopy the impact of Myc deletion. This study thus provides a comprehensive map of how Myc selectively shapes T cell phenotypes and reveals that Myc induction of amino acid transport is pivotal for subsequent bioenergetic and biosynthetic programs.
Project description:RRP41 is one of the nine subunits constituting the core complex of the eukaryotic RNA exosome. Data presented here allowed a proteomic characterization of the interactome of wild type or mutated myc-tagged RRP41 proteins incorporated into the RNA exosome. Flowers from myc-tagged RRP41 expressing lines were used to affinity purify RNA exosomes, which were analyzed by nanoLC-MS/MS analysis. Non-transgenic Arabidopsis lines of Col-0 accession were used in mock-immunoprecipitation experiments as negative controls.
Project description:We perfomed the transcriptomic analysis of differential genetic expression in the CosR-knockdown condition with a DNA microarray. CosR-specific antisense-PNA was used for CosR-knockdown. The results revealed that CosR significantly affected the expression of several genes involved in various cellular function in Campylobacter. Final goal of this study is to figure out the importance and the role of CosR in Campylobacter by identification of the CosR regulons. Two-condition experiment, WT vs. CosR-knockdown. Biological replicates: 3 wild type (control) replicates, 3 CosR-knockdown replicates. For preparing the total RNA, C. jejuni cells were grown at 42M-BM-0C in MH broth supplemented with 1.5 M-NM-<M CosR-PNA for CosR knockdown to the mid-exponential phase for approximately 8 hr with shaking.