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Kinetic analysis of TGFbeta-induced EMT in NMuMG/E9 cells

ABSTRACT: Kinetic analysis of TGFbeta-induced EMT in NMuMG/E9 cells

PROVIDER: PRJNA449023 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR6954260.fastq.gz	Fastqsanger.gz
	SRR6954270.fastq.gz	Fastqsanger.gz
	SRR6954280.fastq.gz	Fastqsanger.gz
	SRR6954290.fastq.gz	Fastqsanger.gz
	SRR6954300.fastq.gz	Fastqsanger.gz

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Kinetic analysis of TGFbeta-induced EMT in NMuMG/E9 cells

Project description:To investigate the transcriptional remodelling during EMT, we treated normal murine mammary gland epithelial cells with TGFbeta for 0, 2h, 6h, 12h, 24h, 36h, 48h, 60h, 72h, 96h, 168h and 240h. Using WGCNA and pathway enrichment analysis we identified multiple gene expression modules that were enriched in general, signaling, metabolic or stuctural pathways highly relevant for EMT.

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The dual role of Irf1 in maintaining epithelial identity while enabling EMT [ChIP-seq]

Project description:To investigate the context-dependent function of Irf1 in maintaining epithelial identity while enabling TGFbeta-induced EMT in NMuMG/E9 cells, we performed chromatin immunoprecipitation with Irf1-specific antibodies in NMuMG cells treated for 2 days with TGFbeta or left untreated (0d TGFbeta). Intersection with RNA-sequencing after downregulation of Irf1 with or without treatment with TGFbeta for two days revealed genes that are directly regulated by Irf1 and that could contribute to the dual role of Irf1 in EMT.

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Mesenchymal Epithelial Transition (MET) Timecourse of NMuMG-E9 cells treated with TGFb

Project description:Background: Epithelial-mesenchymal transition (EMT) has been implicated in metastasis, drug resistance, survival under stress and also conferring stem cell-like traits to cancer cells. However, several of the studies have been carried out using model systems that don’t appropriately recapitulate all stages of the dynamic process of EMT. Hence, there is a need to overcome this limitation by development of a model system that allows us to mimic each stage of EMT and accurately assess the plastic changes associated with it. Methods: We have derived a cancer cell line from the PyMT-MMTV model of breast cancer, named PyMT-1099 cells, and undertaken a detailed characterization of the morpho-genetic changes it undergoes during a TGFbeta-induced EMT. Further, we have also performed high throughput transcriptomics on PyMT-1099 cells undergoing EMT in a high resolution kinetic of TGFbeta treatment. Results: We show that PyMT-1099 cells undergo an EMT comparable to the classically used immortalized NMuMG cells as assessed by morphological, and marker expression changes on TGFbeta treatment. Further, PyMT-1099 cells can also migrate in vitro in response to TGFbeta treatment. These cells are also tumorigenic and lead to metastasis formation when transplanted into immunocompromised mice. Conclusion: In this study we report the development of PyMT-1099 cells as an excellent tool to model and study breast cancer-associated EMT both in vitro and in vivo and show that these cells overcome the limitations posed by other cellular systems currently being used to study EMT

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Project description:To investigate the context-dependent function of Irf1 in maintaining epithelial identity while enabling TGFbeta-induced EMT in NMuMG/E9 cells, we downregulated Irf1 by siRNA and analyzed differentially regulated genes and pathways upon EMT induction (2 days TGFbeta) or in the absence of EMT (0 day TGFbeta). Intersection with Irf1 ChIP-sequencing after 2 days of TGFbeta treatment or in untreated cells revealed genes that are directly regulated by Irf1 and that could contribute to the dual role of Irf1 in EMT.

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Foxf2 plays a dual role during TGFb-induced EMT by promoting apoptosis yet enabling cell junction dissolution and migration.

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Mesenchymal Epithelial Transition (MET) Timecourse of NMuMG-E9 cells treated with TGFb

Project description:Mesenchymal Epithelial Transition (MET) Timecourse of NMuMG-E9 cells treated with TGFb

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Gene expression from NMuMG cells overexpressing major satellite treated with TGFbeta

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Project description:We report the gene expression in mouse NMuMG cells with or without UGCG depletion using RNA sequencing. After data analysis, the 11218 transcripts were identified. Among these transcripts, the TGF-β signaling target genes and EMT marker genes were significantly upregulated in UGCG depleted NMuMG cells. Then we used these genes and mouse TGF-β or EMT gene signatures to do the gene set enrichment analysis and showed the negative correlation of UGCG and TGF-β signaling or EMT in NMuMG cells.

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TGF-β and RREB1 coordinate EMT programs in NMuMG cells

Project description:TGF-β and RREB1 coordinate EMT programs in NMuMG cells

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Genome-wide transcripts in NMuMG cells expressing Osr2

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