Project description:Chemically reprogramming somatic cells to iPSCs is time-consuming and low-efficiency. Here, we discribe a rapid chemical reprogramming condition enabling generating iPSCs from MEFs in 12 days. To further investigate the mechemisms and draw an epigenetic maps during the rapid chemical reprogramming, we performed time-course RNA-seq, ATAC-seq, RRBS and CUT&Tag (H3K4me3, H3K18la, H3K9me3, H3K27me3 and H3K27ac).
Project description:To reveal the role of NCOA7 in cellular senescence, we performed the CUT&TAG assay using the H3K27ac antibody to map acetylation in granulosa cells from control and POI patients. We conducted the DNA sequencing of libraries from CUT&TAG assay using the H3K27ac antibody in human primary granulosa cells.
Project description:To determine the molecular regulation of Tregs by Setd2, large intestine Tregs (CD3+CD4+Foxp3-YFP+) from Setd2-deficient and control Tregs were subjected to H3K27ac CUT&Tag analysis.
Project description:Cleavave Under Targets and Tagmentation (CUT&TAG) sequencing for YAP, H3K27ac, and H3K27me3 of tongue epithelia-derived, transgene-selected primarily cultured cells from control mice or transgenic mice expressing HPV16E6-E7 and YAP1S127A
Project description:We show that CD-RXRα axis significantly promotes mouse chemical reprogramming of fibroblasts. To identify the molecular mechanisms of CD-RXRα axis in chemical reprogramming, we construct Flag-Rxra-OE cells. We performed Flag-Rxra CUT&Tag with reprogramming intermediate, also we performed CUT&Tag with or without CD treatment for reprogramming intermediates.
Project description:CUT& Tag was performed using H3K27ac antibody to identify the super-enhancers of GC with different growth patterns through ROSE algorithm. Comparison of SE-driven oncogenes in infiltrative and expanding GC was carried out to screen key signatures and candidate genes.
Project description:In this study, we found that H3K23ac, H3K27ac, and H4K5ac level are elevated in VSMCs of aneurysmal aortas. To further explore the potential functional significance of H3K23ac, H3K27ac, and H4K5ac in aortic aneurysm, we performed genome-wide cleavage under targets and tagmentation (CUT&Tag) analysis to identify candidate genes regulated by H3K23ac, H3K27ac, or H4K5ac in rat VSMC. Following CUT&Tag, H3K23ac, H3K27ac, or H4K5ac-associated DNAs were amplified using non-biased conditions, labeled, and sequenced with Illumina NovaSeq 150PE.
Project description:To gain mechanistic insight into how Epigenetic factors reprogramming tumor immune microenvironment in response to IFNγ stimulation. we performed CUT&Tag sequencing in murine PDAC cells.For comparison of the transcription profile between IFNγ-PD-L1all and IFNγ+PD-L1all cells, we treated KPC1199 cells with vehicle control or 10 ng/mL recombinant mouse IFNγ for 20 hours and harvested them. For comparison of PD-L1hi with PD-L1all, cells were either sorted with the top 15% PD-L1+ or total of IFNγ+PD-L1all PDAC cells, and freshly collected as soon as possible. Hyperactive pA-Tn5 Transposase for CUT&Tag kit from Vazyme (TD901), and antibodies against H3K4me3 from Abcam (ab213224), H3K27Ac from Abcam (ab4729),H2A119Ub from Cell Signaling (8240T) and H3K27me3 from Abcam (ab6002) were employed. Trueprep index kit v2 and v3 for illumina were used to establish DNA library.
Project description:Purpose: To transcriptome widely reveal histone acetylation modification H3K9ac and H3K27ac regulation by Rbm25, we performed Cleavage Under Targets and Tagmentation (CUT&Tag) analysis of wild-type (WT) and Rbm25-deficiency BMDMs. Methods: Mouse BMDMs were generated from bone marrow cells in RPMI-1640 medium with recombinant mouse M-CSF (20ng/ml). BMDMs were stained to confirm the surface expression of CD11b and F4/80. Cells with purity >97.5% were used for subsequent experiments. WT and Rbm25 deficient BMDMs were stimulated with LPS (100ng/ml) for 4 hours. Results: We carried out CUT&Tag assays using antibody against H3K9ac or H3K27ac, and found the global influence of Rbm25 in regulating H3K9ac and H3K27ac of transcription regulatory regions of key genes for inflammatory macrophage effector function.
