Effect of Eya3 knockdown and constitutively-active NF-kB add-back (IKK2-SE) in 66cl4 cells in vitro
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE288294 | GEO | 2025/05/14
REPOSITORIES: GEO
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Project description:Effect of Eya3 knockdown and constitutively-active NF-kB add-back (IKK2-SE) in 66cl4 cells in vitro
| PRJNA1216787 | ENA
Project description:To investigate gene expression changes associated with Eya3 in the 66cl4 cell line
2025-05-14 | GSE288293 | GEO
Project description:The important function of Eya (eyes absent) in eye development of D.melanogaster is well known, the function of its mammalian orthologs Eya1-4 is only partially understood. Yet, no phenotype for Eya2 and Eya3 in humans or mouse have been described. In our study we analysed new Eya-deficient mouse model generated by insertional mutagenesis. Expression analysis of Eya3 by in-situ hybridizations and M-NM-2-Gal-staining revealed a broad expression of the gene throughout development, e.g. in brain, eyes, branchial arches, somites and limbs. This diverse expression pattern suggests pleiotropic effects of Eya3 in mammalian organisms. In contrast to D. melanogaster, a close examination of the eye by various methods did not show any obvious ocular defects in mutant animals. A broad phenotypical characterization of homozygous mutants displayed decreased body weight and body length. Additionally, behavioral changes as a educed forward locomotor activity and altered exploratory behaviour were identified. A significant reduction of forelimb grip strength and an impairment of cardiac processes in Eya3-/- mice suggest a role for Eya3 in muscle development and heart function. Our results implicate that Eya3 plays various roles outside the eye with effects particularly on heart and muscle function in the mouse. Keywords: Eya3-deficient mice, cDNA microarrays, brain, heart, muscle analysis of 4 Eya3 mutant mice versus a pool of 5 wildtype animals; brain, heart, and muscle; 8 hybridisations for each tissue: two technical replicates for each mutant animal including one dye-swap experiments
2008-12-17 | E-GEOD-9999 | biostudies-arrayexpress
Project description:Inflammation is a major factor in heart disease. IκB kinase (IKK) and its downstream target NF-κB are regulators of inflammation and are activated in cardiac disorders, but their precise contributions and targets are unclear. We analyzed IKK/NF-κB function in the heart by a gain-of-function approach, generating an inducible transgenic mouse model with cardiomyocyte-specific expression of constitutively active IKK2 (IKK2-CA). In adult animals, IKK2 activation led to inflammatory dilated cardiomyopathy and heart failure. Transgenic hearts showed infiltration with CD11b+ cells, fibrosis, fetal reprogramming, and atrophy of myocytes with strong constitutively active IKK2 expression. To gain insight into proximal events after activation of IKK2/NF-κB, we isolated cardiomyocytes and activated the transgene in vitro, thus avoiding interference of other cell types. Gene expression analysis revealed up-regulation of transcripts of chemotactic cytokines, adhesion molecules, and a striking number of IFN-regulated genes in cardiomyocytes expressing IKK2-CA. In addition, apoptotic and anti-apoptotic genes were up-regulated.
2015-10-19 | E-MTAB-3975 | biostudies-arrayexpress
Project description:The important function of Eya (eyes absent) in eye development of D.melanogaster is well known, the function of its mammalian orthologs Eya1-4 is only partially understood. Yet, no phenotype for Eya2 and Eya3 in humans or mouse have been described. In our study we analysed new Eya-deficient mouse model generated by insertional mutagenesis. Expression analysis of Eya3 by in-situ hybridizations and β-Gal-staining revealed a broad expression of the gene throughout development, e.g. in brain, eyes, branchial arches, somites and limbs. This diverse expression pattern suggests pleiotropic effects of Eya3 in mammalian organisms. In contrast to D. melanogaster, a close examination of the eye by various methods did not show any obvious ocular defects in mutant animals. A broad phenotypical characterization of homozygous mutants displayed decreased body weight and body length. Additionally, behavioral changes as a educed forward locomotor activity and altered exploratory behaviour were identified. A significant reduction of forelimb grip strength and an impairment of cardiac processes in Eya3-/- mice suggest a role for Eya3 in muscle development and heart function. Our results implicate that Eya3 plays various roles outside the eye with effects particularly on heart and muscle function in the mouse. Keywords: Eya3-deficient mice, cDNA microarrays, brain, heart, muscle
2008-12-17 | GSE9999 | GEO
Project description:Effect of shRNA-mediated knockdown of Eya3 in 66cl4 murine TNBC cells in vitro
| PRJNA1216788 | ENA
Project description:Eyes absent homolog 3 (EYA3) is a transcriptional cofactor that is known to partner with the sine oculis homeobox (SIX) family to regulate gene expression. In this study, we characterized a novel splice isoform of EYA3, including the alternative cassette exon 7 (EYA3+ex7), that becomes more highly expressed compared to the isoform lacking exon 7 (EYA3Δex7) during muscle cell differentiation and muscle development. This developmentally-regulated splicing pattern was evolutionarily conserved in humans, mice, rats, chickens, and zebrafish. We found that EYA3 isoforms are vital to proliferation and differentiation of myoblasts in a cell culture model of myogenesis. To understand the underlying mechanism, we defined the transcriptional networks controlled by EYA3 through RNA-sequencing of EYA3 depleted cells. We then hypothesized that binding partners of EYA3+ex7 and EYA3Δex7 might contribute to the mechanism by which they regulate transcription. An affinity-purification mass spectrometry approach was used to identify binding partners of EYA3 isoforms by heterologous expression of hemagglutinin (HA) tagged isoforms in HEK293T cells. 26 binding partners (defined as those with log2(FC) > 1 and p < 0.05 compared to HA epitope tag control) were shared among the EYA3 isoforms, including five transcription factors: zinc finger and BTB domain containing 1 (ZBTB1), tumor protein p53 (TP53), SIX5, SIX2, and SIX4. We next demonstrated that ZBTB1 and SIX4 share overlapping transcriptional targets with EYA3, indicating that EYA3 partners with ZBTB1 or SIX4 to regulate gene expression. Three and thirteen unique binding partners were discovered for EYA3Δex7 and EYA3+ex7, respectively. Therefore, inclusion of Eya3 exon 7 may allow for unique interactions and contribute to functions of EYA3+ex7 that are distinct from EYA3Δex7. This dataset includes affinity purified data for Eya3 isoforms in HEK293T cells.
2023-10-13 | PXD037572 | Pride
Project description:Hyperactivation of JAK2 kinase is a unifying feature of human Ph- myeloproliferative neoplasms (MPNs), most commonly due to the JAK2 V617F mutation. Mice harboring a homologous mutation in the Jak2 locus exhibit a phenotype resembling polycythemia vera. NFκB pathway hyperactivation is present in myeloid neoplasms, including MPNs, despite scarcity of mutations in NFκB pathway genes. To determine the impact of NFκB pathway hyperactivation in conjunction with Jak2 V617F, we utilized Ikk2 (Ikk2-CA) mice. Pan-hematopoietic Ikk2-CA alone produced depletion of hematopoietic stem cells and B cells. When combined with the Jak2 V617F mutation, Ikk2-CA rescued the polycythemia vera phenotype of Jak2 V617F. Likewise, Jak2 V617F ameliorated defects in hematopoiesis produced by Ikk2-CA. Single-cell RNA sequencing of hematopoietic stem and progenitor cells revealed multiple genes antagonistically regulated by Jak2 and Ikk2, including subsets whose expression was altered by Jak2 V617F and/or Ikk2-CA but partly or fully rectified in the double mutant. We hypothesize that Jak2 promotes hematopoietic stem cell population self-renewal, whereas Ikk2 promotes myeloid lineage differentiation, and biases cell fates at several branch points in hematopoiesis. Jak2 and Ikk2 both regulate multiple genes affecting myeloid maturation and cell death. Therefore, the presence of dual Jak2 and NFκB hyperactivation may present neomorphic therapeutic vulnerabilities in myeloid neoplasms.
2025-04-07 | GSE263101 | GEO
Project description:The transcription factor NF-κB has been associated with a range of pathological conditions of the heart, mainly based on its function as a master regulator of inflammation and pro-survival factor. Here, we addressed the question what effects activation of NF-κB can have during murine heart development. We expressed a constitutively active (CA) mutant of IKK2, the kinase activating canonical NF-κB signaling, specifically in cardiomyocytes under the control of the α-myosin heavy chain promoter. Expression of IKK2-CA resulted in embryonic lethality around E13. Embryos showed defects in compact zone formation and the contractile apparatus, and overall were characterized by widespread inflammation with infiltration of myeloid cells. Gene expression analysis at E12.5 suggested an interferon type I signature, with increased expression of interferon regulatory factors.
2015-10-14 | E-MTAB-3971 | biostudies-arrayexpress
Project description:Native MS analysis of prokaryotically expressed and purified EYA3. Native MS analysis of EYA3 phosphorylation by Src and EYA3 autodephosphorylation reaction.
2019-12-17 | PXD016596 | Pride