![]() Inhibition of autophagy determined by the detection of LC3-II expression. Expression of mitochondrial matrix proteins including HSP60 and COX4 were by Western blotting. PK-15 and 3D4/2 cells infected with CSFV (MOI = 1) in the presence or absence of 3-MA (5 mM) at 48 hpi. (C) Changes of mitochondrial proteins in CSFV-infected PK-15 and 3D4/2 cells treated with 3-MA. (B) Changes of mitochondrial proteins in CSFV-infected 3D4/2 cells were analyzed as in (A). P values were calculated using two-way ANOVA. The histograms on the right showed the statistical analysis of the protein band intensity (mean ± SD n = 3 *P < 0.05 ** P< 0.01 *** P < 0.001). GAPDH was used as an internal loading control. CSFV infection was verified by immunoblotting with anti-CSFV Npro antibody. The expression of mitochondrial proteins, including MFN2, HSP60, VDAC1, TOM20 and COX4, were analyzed by Western blotting. PK-15 cells were mock-infected or infected with CSFV (MOI = 1), and whole cell lysates (WCL) were prepared at 24, 36 and 48 hpi. (A) Changes of mitochondrial proteins in CSFV-infected PK-15 cells. The decline in number of mitochondria in CSFV-infected cells was related to mitophagy. Our data for mitochondrial fission and selective mitophagy in CSFV-infected cells reveal a unique view of the pathogenesis of CSFV infection and provide new avenues for the development of antiviral strategies. The preservation of mitochondrial proteins, upregulated apoptotic signals and decline of viral replication resulting from the silencing of Drp1 and Parkin in CSFV-infected cells suggested that CSFV induced mitochondrial fission and mitophagy to enhance cell survival and viral persistence. Mitochondrial fission caused by CSFV infection was further determined by mitochondrial fragmentation and Drp1 translocation into mitochondria using a confocal microscope. In addition, a sensitive dual fluorescence reporter (mito-mRFP-EGFP) was utilized to analyze the delivery of mitophagosomes to lysosomes. Mitophagosomes and mitophagolysosomes induced by CSFV were, respectively, observed by the colocalization of LC3-associated mitochondria with Parkin or lysosomes. Upon activation of the PINK1 and Parkin pathways, Mitofusin 2 (MFN2), a mitochondrial fusion mediator, was ubiquitinated and degraded in CSFV-infected cells. CSFV infection increased the expression and mitochondrial translocation of Pink and Parkin. The formation of mitophagosomes and decline in mitochondrial mass relevant to mitophagy were detected in CSFV-infected cells. In this study, we offer the first evidence that CSFV induces mitochondrial fission and mitophagy to inhibit host cell apoptosis for persistent infection. The balance of mitochondrial dynamics is essential for cellular homeostasis. Oxidative stress and a reduced mitochondrial transmembrane potential are disturbed in CSFV-infected cells. Novel Global Community Educational Foundation,AustraliaĪbstract: Objective: The dramatic increase in the population with dementia expected in the nextClassical swine fever virus (CSFV), which causes typical clinical characteristics in piglets, including hemorrhagic syndrome and immunosuppression, is linked to hepatitis C and dengue virus.Title:Biomarkers for Alzheimer's Disease DiagnosisĪuthor(s): Vasileios Mantzavinos and Athanasios Alexiou* Mitochondrial dynamics, mild cognitive impairment. Keywords: Alzheimer's disease biomarkers, oxidative stress, metal ions, vascular disorders, protein dysfunctions, Vascular disorders, protein dysfunctions and alterations in the mitochondrial populations.Ĭonclusion: A multiparametric model of Alzheimer's biomarkers is presented according to the latest Several potential markers are discussed such as oxidative stress, metal ions, The aim of this study is to investigate the main risk factors thatĪffect and increase Alzheimer's disease progression over time even in cases with no significant memory While Alzheimer's disease is the most commonĬause of dementia, by the time it is typically diagnosed, substantial neuronal loss and neuropathological Objective: The dramatic increase in the population with dementia expected in the nextĭecades is accompanied by the establishment of novel and innovated methods that will offer accurateĪnd efficient detection of the disease in its early stages.
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