Project description:Analysis of STAT1 and IRF1 binding in IFNg treated and untreated HeLa cells for 6 hours was done using 50mer oligonucleotide probes at 30bp intervals tiling over non-repetitive 16MB gene locus (HG17).
Project description:Analysis of H3ac, H4ac, STAT1 and IRF1 binding in IFNg treated and untreated HeLa cells for 6 hours was done using 50mer oligonucletide probes at 30bp intervals tiling over non-repetitive 122kb CIITA locus(HG17) Keywords: ChIP-chip
Project description:Analysis of H3ac, H4ac, STAT1 and IRF1 binding in IFNg treated and untreated HeLa cells for 6 hours was done using 50mer oligonucletide probes at 30bp intervals tiling over non-repetitive 16MB gene locus(HG17) Keywords: ChIP-chip
Project description:Analysis of H3ac, H4ac, STAT1 and IRF1 binding in IFNg treated and untreated HeLa cells for 6 hours was done using 50mer oligonucletide probes at 30bp intervals tiling over non-repetitive 16MB gene locus(HG17) Keywords: ChIP-chip three replicates for each marker at each state.
Project description:STAT1 ChIP-chip performed on Human Hela S3 Cells for three different platforms. Nimblegen ENCODE arrays which comprise 50mer oligonucleotides spaces every 38bps (overlapping by 12nts) (5 biological replicates), custom maskless array tiling most of ENCODE with 50mer oligonucleotides end-to-end (3 biological replicates) and custom maskless array tiling most of ENCODE with 36mer oligonucleotides end-to-end (2 biological replicates). The chromatin-immunoprecipitation protocol is the same for all samples, however the labelling and hybridization protocols differ between Nimblegen and custom maskless arrays. Keywords = Transcription Factor Binding, STAT1, ChIP-chip, Human, Genome Tiling Arrays Keywords: other
Project description:DUX4 activates the first wave of zygotic gene expression in the early embryo. Mis-expression of DUX4 in skeletal muscle causes facioscapulohumeral dystrophy (FSHD), whereas expression in cancers suppresses IFNg-induction of MHC Class I and contributes to immune evasion. We show that the DUX4 protein broadly suppresses immune signaling pathways—including IFNg, IFNb, DDX58, IFIH1 and cGAS mediated pathways. A conserved region containing (L)LxxL(L) motifs in the DUX4 carboxyterminal domain (CTD) was necessary to suppress interferon stimulated genes (ISGs). Co-immunoprecipitation identified DUX4-CTD interaction with multiple immune signaling factors, including STAT1. The DUX4-CTD (L)LxxL(L) region interacts with phosphorylated STAT1, sequesters it in the nucleus, modestly reduces its DNA binding, and prevents STAT1 from inducing ISG transcription. Mouse Dux similarly interacted with STAT1 and suppressed IFNg induction of ISGs. These findings identify an evolved role of the DUXC family in modulating immune signaling pathways with implications for development, cancers, and FSHD.
Project description:STAT1 is the major transcription factor (TF) driving response to exposure to IFNg. C-JUN is a TF which has been shown to bind with STAT1 and act as a partner TF. We have expression profiled WT and CJUN-/- MEFs in order to determine the set of IFNg genes regulated by STAT1 and C-JUN. This study was designed primarily to test the accuracy of in silico cis-regulatory module prediction algorithm called SCRM. The set of genes differentially expressed after IFNg are likely to be targets of the TF STAT1. A subset of these genes will also be targeted by partner TFs of STAT1 (e.g. C-JUN). By measuring the expression of the WT IFNg responsive genes in a C-JUN-/- model, the subset of genes that are regulated by C-JUN can be ascertained. This set of genes can then be compared against those predicted to be regulated by C-JUN using the in silico approach SCRM, and therefore the accuracy of the in silico predictions can be determined. Cells were treated with both IFNg and cycloheximide (CHX) to determine direct target genes of the TF STAT1. CHX was added to reduce the downstream response of IFNg treatment (via inhibition of protein synthesis) , resulting in a high quality list of genes directly targeted by STAT1. CHX treatment also allowed for a longer (3hr rather than 1hr) IFNg treatment time, resulting in a more robust IFNg gene signature. Microarrays were analysed using the aroma package in R and the differentially expressed genes were determined through analysis using the limma package. Genes were ranked based on the FDR adjusted p-value calculated using a t-test between IFNg + CHX treated cells and untreated cells (in both WT and C-JUN-/- MEFs). Significantly upregulated genes (P<0.05) were considered direct targets of STAT1.
Project description:We have identified a number of miRNAs that are differentially expressed in LPS and IFNγ treated RAW264.7 cells, as compared to untreated cells. Microarray and quantitative real-time PCR (qRT-PCR) results showed that miR-103 decreased while the STAT1 expression increased substantially in RAW264.7 derived M1 macrophage, and there was a significant negative correlation between miR-103 and STAT1.Overexpression of miR-103 suppressed M1 polarization by inhibiting STAT1/IRF1 signaling pathway and vice versa in vitro. STAT1 is a direct target of of miR-103.
