Project description:Transcriptional profiling of human hepatoma cell lines comparing control uninfected Huh7.5 cells with Huh7.5 cells persistently infected with HCV (HPI cell). The latter maintains and produces HCV.
Project description:Cytosolic lipid droplets (LDs) are vital to Hepatitis C Virus (HCV) infection as the putative sites of virion assembly. To identify novel regulators of HCV particle production, we performed quantitative LD proteome analysis. Huh7.5 cells were labeled by stable isotope labeling with heavy amino acids in cell culture (SILAC) and subsequently infected with an HCV Jc1 reporter virus. After selection for HCV-infected cells, equal amounts of HCV-infected and uninfected control cells were mixed, LDs were isolated and analyzed by LC-ESI-MS/MS.
Project description:huh7.5 hepatic cell lines were infected with HCV. RNA and Protein were extract after 72 hours and The Human Adipogenesis RTM-BM-2 ProfilerM-bM-^DM-" PCR ArrayM-BM- was used to compare the gene expression in the two group. qPCR gene expression profiling. HCV infected hepatic cells vs uninfected hepatocyte. Equal amount total RNA from each samples were used
Project description:Differential expression study of lncRNAs (long non-coding RNAs) and mRNAs in HCV infection. For this we infected Huh7.5 cells with HCV (JFH-1) virus and control cells were mock infected. 48h post infection we isolated total RNA using qiagen RNeasy kit as per manufacturer's protocol. The total RNA was then subjected to microarray for both lncRNAs and mRNAs. The resultant profile can be used to compare gene expression in HCV infection versus mock infected cells.
Project description:TMT10plex global proteomics of HCV infected HuH7.5 cells or HCV infected humanized mouse livers. TMT channels were combined after labeling and separated off line with a highPH reverse phase fractionation into 24 fractions.
Project description:Huh7.5 cells were infected in duplicate with 1b Con1cc virus. At peak infection (>80% cells were HCV positive, day 7 post infection), total RNA was isolated using TRIzol LS reagent. Analysis of the transcriptomes of Huh7.5 cells disturbed by Con1cc infection. We found that in addition to activation of apoptosis and antiviral response in the cell, Con1cc infection also induced angiogenesis-related genes and disturbed cholesterol metabolism. This study provided experimental evidence towards the understanding of molecular mechanisms of HCV pathogenesis.
Project description:huh7.5 hepatic cell lines were infected with HCV. RNA and Protein were extract after 72 hours and The Human Adipogenesis RT² Profiler™ PCR Array was used to compare the gene expression in the two group.
Project description:Drugs directly targeting Hepatitis C (HCV) are often rendered useless by the high mutation rate of the virus. Thus, we deduce that targeting of host factor that affect HCV replication may provide enhanced therapy fort HCV infection. Hepatocyte cell line Huh7 is known to be non-permissive for Hepatits C (HCV) replication. Through a method developed by the Rice laboratory (Blight, K.J., et al., J Virol, 2002), selection of a small subset of permissive hepatocytes is possible. The Rice laboratory generated the first permissive cell line, Huh7.5, using this method. We generated another permissive cell line, HRP1, using the same method. With microarray, we compared the expression of host mRNAs in non-permissive Huh7 to both Huh7.5 and HRP1 searching for host factors lost in the cell lines permisive for HCV replication. Non-permissive cell line Huh7 and permissive cell lines Huh7.5 and HRP1 were harvested for RNA extraction and hybridization on Affymetrix microarrays.
