Project description:To investigate the genome-wide transcription targets of GR in renal cell carcinoma, cleavage under targets and tagmentation (CUT&Tag) was performed in Caki-1 cells using antibodies againstGR. Following CUT&Tag, DNAs were amplified using non-biased conditions, labeled, and sequenced with Illumina NovaSeq 150PE.
Project description:To investigate the genome-wide transcription targets of H4K12 lactylation in ccRCC cells, cleavage under targets and tagmentation (CUT&Tag) was performed in 786-O and Caki-1 cells using antibody against H4K12 lactylation. Following CUT&Tag, DNAs were amplified using non-biased conditions, labeled, and sequenced with Illumina NovaSeq 150PE.
Project description:To investigate the genome-wide transcriptional targets associated with H4K12 lactylation in renal cell carcinoma, cleavage under targets and tagmentation (CUT&Tag) was performed in 786-O cells with or without dexamethasone treatment using specific antibodies against H4K12 lactylation or GR. Following CUT&Tag, genomic DNA fragments were amplified under non-biased conditions and sequenced on the Illumina NovaSeq 150PE platform to generate high-resolution maps of H4K12 lactylation or GR–associated chromatin regions.
Project description:Genome-wide identification of transcriptional targets of H4K12 lactylation and GR in 786-O cells with or without dexamethasone treatment.
Project description:Comparison of various inductibles constructions of arabidopsis treated by differents solutions. The aim of this project is to determine the primary and secondary targets of the LEC2 transcrition factor. The induction by the dexamethasone allows the expression of constructions LEC2:GR or ttg:GR introduced into transforming plants.The dexamethasone induces the expression of the contruct which carries the receptor to the glucocorticoïde (GR.). The cycloheximide is a inhibitor of the translation and allows, coupled with the dexamethasome, the identification of the primary targets only. The LEC2_GR25 and LEC2_GR31 constructs are two independents transforming plants of the cDNA-LEC2_GR construct. Both were used in order to have an idea of the variability of the data transcriptome obtained. The ttg_GR construct is used here as control ; ttg is another transcription factor, thus if induction with the dexamethasone works well, the targets induced in the two mutants will be different.
Project description:To compare the transcriptomic changes mediated by VHL knockout in Caki-1 cells, we carried out the next-generation sequencing (NGS)-derived transcriptomic profiling. We found significant enrichment of gene signatures involving IFNa/b signaling in VHL-KO Caki-1 cells compared to control cells. Our analysis also demonstrated that VHL deficiency induced the expression of genes associated with the cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) pathway involved in sensing the cytoplasmic presence of double-stranded DNA (dsDNA).
Project description:Exogenous glucocorticoids are widely used in the clinic for the treatment of inflammatory disorders and auto-immune diseases. Unfortunately, their use is hampered by many side effects and therapy resistance. Efforts to find more selective glucocorticoid receptor (GR) agonists and modulators (called SEGRAMs), able to separate anti-inflammatory effects via gene suppression from metabolic effects via gene activation, have been unsuccessful so far. In this study, we characterized a set of functionally diverse GR ligands in A549 cells, first using a panel of luciferase-based reporter gene assays evaluating GR-driven gene activation and gene suppression. We expanded this minimal assay set with novel luciferase-based read-outs monitoring GR protein levels, GR dimerization and GR Serine 211 (Ser211) phosphorylation status and compared their outcomes with compound effects on the mRNA levels of known GR target genes in A549 cells and primary hepatocytes. We found that luciferase reporters evaluating GR-driven gene activation and gene repression were not always reliable predictors for effects on endogenous target genes. Remarkably, our novel assay monitoring GR Ser211 phosphorylation levels proved to be the most reliable predictor for compound effects on almost all tested endogenous GR targets, both driven by gene activation and repression. The integration of this novel assay in existing screening platforms may therefore increase chances to find novel GR ligands with an improved therapeutic benefit.