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Transcriptome analysis among senescent cells induced by various stimuli

ABSTRACT: Transcriptome analysis among senescent cells induced by various stimuli

PROVIDER: PRJNA576221 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR10238700.fastq.gz	Fastqsanger.gz
	SRR10238700_1.fastq.gz	Fastqsanger.gz
	SRR10238700_2.fastq.gz	Fastqsanger.gz
	SRR10238701.fastq.gz	Fastqsanger.gz
	SRR10238701_1.fastq.gz	Fastqsanger.gz

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Transcriptome analysis among senescent cells induced by various stimuli

Project description:We carried out the analysis using human skin fibrobllast HCA2 (MJ90) cells. Cells were indcued to be senescent cells by treatment with doxorubicin for 24 hrs, tert-butyl-hydroperoxide for 24 hrs, or serial passages as well as nultin3a + RO3306. As a result, we found simailar transcriptome signatures among senescent cells induced by various stimuli

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NET induced by different stimuli: a comparative proteomic analysis

Project description:Neutrophil extracellular traps (NET) formation is part of the neutrophil response to infections, but excessive or inappropriate NETosis may trigger the production of autoantibodies and cause organ damage in autoimmune disorders. Spontaneously netting neutrophils are not frequent and induction of NET in vitro by selected stimuli is necessary to investigate their structure. In the present work, the protein composition and post-translational modifications of NET produced under different stimuli is studied by means of proteomic analysis. Neutrophils from healthy donors were stimulated by PMA, A23187, E.Coli LPS or untreated; after 3 hours cells were washed, treated with DNase and supernatants collected for mass spectrometry. Data were analyzed by unsupervised hierarchical clustering analyses. We identified proteins contained in NETs of any source or exclusive of one stimulus: LPS-induced and spontaneous NET diverge in protein composition, while PMA- and A23187-induced NET appear more similar. Among the post-translational modifications we examined, methionine sulfoxidation is frequent especially in PMA- and LPS-induced NETs. Myeloperoxidase is the protein more extensively modified. Thus, proteomic analysis indicates that NETs induced by different stimuli are heterogeneous in terms of both protein composition and post-translational modifications, suggesting that NET induced in different conditions may have different biological effects.

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Project description:We expressed either a wt or a phosphomutant version of ZFP36L1 in IMR90 ER:RAS cells. 7 days upon RAS induction (when the cells reach a fully senescent phenotype) we collected the RNA. ZFP36L1 is a RNA binding protein that binds to AU-rich elements in the 3âUTR of mRNAs and triggers their degradation. Our previous experiments showed that the activity of ZFP36L1 was key in the regulation of the senescent phenotype.By performing RNAseq we have uncovered the effect of expressing a constitutively active mutant of ZFP36L1 within the senescent transcriptome. 4 samples examined: Non-senescent cells (EV - 4OHT), Senescent cells (EV + 4OHT), Senescent cells expressing ZFP36L1wt and Senescent cells expressing ZFP36L1mut

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