Project description:The Wnt pathway is a key regulator of embryonic development, cell growth, differentiation, polarity formation, neural development, carcinogenesis, and stem cell self-renewal, and deregulation of the Wnt signalling is associated with many human disease. The central player in the Wnt pathway is β-catenin, A recent study has shown that β-catenin/Tcf/Lef signaling pathway is an essential growth-regulatory pathway in cardiomyocytes. We used DNA microarrays to detail the global trends in gene expression underlying β-catenin-overexpressed cardiomyocytes and identified distinct classes of up- or down-regulated genes during this process. Our findings suggest that β-catenin plays a critical role in regulating cardiac dysfunction at transcriptional level and may provide novel insight into how β-catenin modulates heart diseases. Cardiomyocytes were infected with GFP control or β-catenin adenoviruses for RNA extraction and hybridization on Affymetrix microarrays. We sought to define the effects of β-catenin on the global programme of gene expression in primary cardiomyocytes. To that end, neonatal rat cardiomyocytes were infected with GFP control (G) or β-catenin adenovirus (B) for 24 hours.
Project description:So far very little is known about the fine regulation of some of these intracellular interactions of β-DG and how they are perturbed in diseases. To start filling this gap, HEK-293 cells were transiently transfected with a plasmid carrying the β-DG subunit with GFP fused at its C-terminus. Immunoprecipitation by anti-GFP antibodies followed by shotgun proteomic analysis to investigate the proteins exclusively matching for β-DG binding, applying the following filters to MS identification data: high confidence, peptide rank 1, peptide length minimum 9 amino acid residues and selecting 2 peptides per protein. A series of already known β-DG interactors have been found, whilst significant new matches, which include potential novel β-DG interactors and their related networks, were identified in diverse subcellular compartments.
Project description:The Wnt pathway is a key regulator of embryonic development, cell growth, differentiation, polarity formation, neural development, carcinogenesis, and stem cell self-renewal, and deregulation of the Wnt signalling is associated with many human disease. The central player in the Wnt pathway is β-catenin, A recent study has shown that β-catenin/Tcf/Lef signaling pathway is an essential growth-regulatory pathway in cardiomyocytes. We used DNA microarrays to detail the global trends in gene expression underlying β-catenin-overexpressed cardiomyocytes and identified distinct classes of up- or down-regulated genes during this process. Our findings suggest that β-catenin plays a critical role in regulating cardiac dysfunction at transcriptional level and may provide novel insight into how β-catenin modulates heart diseases.
Project description:propose: The goals of this study are viral transcript of host cells infected with shaan virus and host cell responses in HEK293, A549 and HRT18 infected with shaan virus. Methods: We infected the virus at an MOI of 0.5 to human tissues-derived cell lines including HEK-293, A549 and HRT18. The infected cells were harvested at 24 hours post infection, and a total RNA was extracted from the harvested cells with Trizol LS reagent. The trimmed reads were mapped to shaan virus genome or GRCh37 human genomic DNA reference. The differentially expressed genes (DEGs) between the virus-infected and mock-infected cells were calculated based on the fold changes of FPKM per each gene. Result: Viral transcripts were expressed differentially depending on the viral genes, it is obvious that the nucleocapsid (N) transcript was most expressed in all infected cells. Gene ontology enrichment analysis based on the DEGs revealed that HEK-293, A549 and HRT18 cells highly expressed the genes related to the antiviral defence system and innate immune response in response to the shaan virus infection. Conclusion: In this study, we describe transcriptome data from the shaan virus-infected cells. The inferred DEGs and their related biological functions can be further investigated to understand the shaanvirus-host interaction.
Project description:A GFP-expressing recombinant A/Puerto Rico/8/1934 influenza virus was used to infect C57BL/6 wild type mice and on day 3 post infection, lung alveolar epithelial cells (AEC) were isolated and sorted based on GFP expression. GFP+ AEC represent the infected AEC and GFP- AEC represent the bystander AEC. AEC were also sorted from uninfected mice to serve as controls.
Project description:GFP or GFP-200 proteins were purified from the corresponding Flp-In™ T REx™ 293 cells using GFP-Trap beads. Purified proteins were analyzed by mass spectrometry.
Project description:To study the heterogeneous activation of Wnt/β-catenin signaling in prostate cancer, we established a Wnt/β-catenin signaling reporting system 7xTCF-EGFP. We then sorted out GFP+ cells and GFP- cells and compared their properties on both molecular and cellular levels.
Project description:We over-expressed biotinylated-thyroid hormone receptor beta 1 (TRb1) in mouse liver using an adenovirus in order to perform ChIP-seq experiments. These microarrays were performed to determine gene expression changes in response to tri-iodothyronine (thyroid hormone; T3) stimulation. A control GFP adenovirus was used and gene expression from these livers was also done as a comparison. We performed microarrays from Ad-GFP-infected propylthiouracil (PTU)-fed livers injected with either saline or T3 and Ad-TRb1-GFP infected livers injected with either saline or T3.