Project description:The miRNA expression profiles of of EBV-infected YT and NK92 cells are compared. YT cells have weaker expression of T-bet and IFNg than NK92 cells. It was found that the EBV-encoded miR-BART20-5p inhibits the expression of T-bet and IFNg in YT cells.
Project description:YT and NK92 cells are EBV-infected lymphoma cells of NK- cell origin. YT cells have weaker expression of T-bet and IFNg than NK92 cells. However, this data set shows that both cell lines have similar mRNA levels, implying post-transcriptional regulations.
Project description:The miRNA expression profiles of of EBV-infected YT and NK92 cells are compared. YT cells have weaker expression of T-bet and IFNg than NK92 cells. It was found that the EBV-encoded miR-BART20-5p inhibits the expression of T-bet and IFNg in YT cells. Approximately 100 ng total RNAs from YT and NK92 cells were used as the input. The GeneChip miRNA 2.0 array from Affymetrix, including probes for 44 EBV-encoded miRNAs and 1105 human miRNAs, was used. The experiments were performed in the microarray core laboratory of National Health Research Institute, Taiwan.
Project description:YT and NK92 cells are EBV-infected lymphoma cells of NK- cell origin. YT cells have weaker expression of T-bet and IFNg than NK92 cells. However, this data set shows that both cell lines have similar mRNA levels, implying post-transcriptional regulations. Affymetrix chips (Human Genome U133 plus 2.0) were used for a genome-wide screen of transcriptional profiles on YT and NK cells. Hybridization and washing of the arrays were done according to the GeneChip® Expression Analysis Technical Manual from Affymetrix. The arrays were scanned with GenePix 4000B (Molecular Devices, Sunnyvale, CA.), and the data were extracted with Affymetrix Microarray suite (MAS) software.
Project description:Gene expression profiling of immortalized human mesenchymal stem cells with hTERT/E6/E7 transfected MSCs. hTERT may change gene expression in MSCs. Goal was to determine the gene expressions of immortalized MSCs.
Project description:Transcriptional profiling of human mesenchymal stem cells comparing normoxic MSCs cells with hypoxic MSCs cells. Hypoxia may inhibit senescence of MSCs during expansion. Goal was to determine the effects of hypoxia on global MSCs gene expression.
Project description:Kynureninase is a member of a large family of catalytically diverse but structurally homologous pyridoxal 5'-phosphate (PLP) dependent enzymes known as the aspartate aminotransferase superfamily or alpha-family. The Homo sapiens and other eukaryotic constitutive kynureninases preferentially catalyze the hydrolytic cleavage of 3-hydroxy-l-kynurenine to produce 3-hydroxyanthranilate and l-alanine, while l-kynurenine is the substrate of many prokaryotic inducible kynureninases. The human enzyme was cloned with an N-terminal hexahistidine tag, expressed, and purified from a bacterial expression system using Ni metal ion affinity chromatography. Kinetic characterization of the recombinant enzyme reveals classic Michaelis-Menten behavior, with a Km of 28.3 +/- 1.9 microM and a specific activity of 1.75 micromol min-1 mg-1 for 3-hydroxy-dl-kynurenine. Crystals of recombinant kynureninase that diffracted to 2.0 A were obtained, and the atomic structure of the PLP-bound holoenzyme was determined by molecular replacement using the Pseudomonas fluorescens kynureninase structure (PDB entry 1qz9) as the phasing model. A structural superposition with the P. fluorescens kynureninase revealed that these two structures resemble the "open" and "closed" conformations of aspartate aminotransferase. The comparison illustrates the dynamic nature of these proteins' small domains and reveals a role for Arg-434 similar to its role in other AAT alpha-family members. Docking of 3-hydroxy-l-kynurenine into the human kynureninase active site suggests that Asn-333 and His-102 are involved in substrate binding and molecular discrimination between inducible and constitutive kynureninase substrates.