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Proteome analysis of Co-IP samples from NCI-H1299 cell nuclear lysate with anti-AGO2 and anti-TNRC6A antibody.

ABSTRACT: Proteome analysis of Co-IP samples from NCI-H1299 cell nuclear lysate with anti-AGO2 and anti-TNRC6A antibody.

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Masahiko Kuroda

PROVIDER: PXD017408 | JPOST Repository | Sat Feb 06 00:00:00 GMT 2021

REPOSITORIES: jPOST

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Dataset's files

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			Action	DRS
	001_exKK0690-1_Control_IgG.raw	Raw
	002_exKK0690-2_AGO2.raw	Raw
	003_exKK0690-3_TNRC6A.raw	Raw
	KK0690_Re-analysis.sdia	Other

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Publications

Nuclear microRNAs release paused Pol II via the DDX21-CDK9 complex.

Ohno Shin-Ichiro SI Oikawa Keiki K Tsurui Toshiaki T Harada Yuichirou Y Ono Kana K Tateishi Mizumo M Mirza Aashiq A Takanashi Masakatsu M Kanekura Kosuke K Nagase Kumiko K Shimada Yoshihisa Y Kudo Yujin Y Ikeda Norihiko N Ochiya Takahiro T Wang Xiaozhong X Kuroda Masahiko M

Cell reports 20220401 2

RNA activation (RNAa) is an uncharacterized mechanism of transcriptional activation mediated by small RNAs, such as microRNAs (miRNAs). A critical issue in RNAa research is that it is difficult to distinguish between changes in gene expression caused indirectly by post-transcriptional regulation and direct induction of gene expression by RNAa. Therefore, in this study, we seek to identify a key factor involved in RNAa, using the induction of ZMYND10 by miR-34a as a system to evaluate RNAa. We id ...[more]

PMID: 35417682

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Identification of miRNA targets in breast cancer cells (Ago2-IP)

Project description:miRNAs regulate mRNA stability and translation through the action of the RNAi-induced silencing complex. In this study, we systematically identified endogenous miRNA target genes by using AGO2 immunoprecipitation (AGO2-IP) and microarray analyses in two breast cancer cell lines, MCF7 and MDA-MB-231, representing luminal and basal-like breast cancer, respectively. The expression levels of ~70% of the AGO2-IP mRNAs were increased by DROSHA or DICER1 knockdown. In addition, integrated analysis of miRNA expression profiles, mRNA-AGO2 interaction, and the 3'-UTR of mRNAs revealed that >60% of the AGO2-IP mRNAs were putative targets of the fifty most abundantly expressed miRNAs. To identify mRNAs associated with AGO2, cell lysate was precleaned with control IgG and immunoprecipitated with anti-human Ago2 (Clone 2E12-1C9, Abnova). Total RNA from cell lysate or coimmunoprecipitated with AGO2 was extracted with Trizol and subjected to microarray analysis, with three biological repeats for each experimental condition.

2013-10-21 | E-GEOD-48158 | biostudies-arrayexpress

polysome-IP profiling experiment : Polysomes of untreated or tunicamycin-treated AC16 cells were immunoprecipitated with anti-MRPS15 antibody, followed by RNA sequencing

Project description:Polysomes of untreated (NT) or tunicamycin-treated (TM) human cardiomyocyte AC16 cells were immunoprecipitated (IP MRPS15) with anti-MRPS15 antibody, followed by RNA sequencing. As a control, RNAseq was performed on polysome-associated RNAs of untreated or tunicamycin-treated human cardiomyocyte AC16 cells before immunoprecipitation (input).

2024-04-01 | E-MTAB-13915 | biostudies-arrayexpress

Colorectal cancer anti-ORF1p LFQ IP-MS

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2019-08-23 | PXD013743 | Pride

Transcriptome expression changes in human gastric epithelial cells co-cultured with Helicobacter pylori lysate of different generations.

Project description:With the colonization of Helicobacter Pylori (H.pylori) in the human stomach, a large amount of Helicobacter Pylori (H.pylori)colonized into the stomach disintegrates and releases a large number plenty of toxic proteins and other bacterial structure components. TollToll-like receptors (TLRs) in gastric epithelial cells which sensitive to the components of bacteria lysate around the microenvironment attach them and activite innate immune and pro-inflammatory response once contact with antigen. The consequences after long-term stimulation of H.pylori lysate in gastric epithelial cells has not been reported . To investigate the effects of H.pylori lysate on TLRs expression and inflammatory cytokines level during long-term stimulation of gastric epithelial cells and clarify the potential mechanismHence, we detected TLRs mRNA level and inflammatory cytokines expression in GES-1 cells co-culture with lysate from 1 to 30 generations. The sensitivity to H.pylori or H.pylori lysate in GES-1 cells were also monitored. We also explored whether LPS was the main factors contributing to this change. Mongolian gerbils were used to replicate H.pylori infection animal model. After transcriptome sequencing，A TLR2/6 heterodimers agonist Pam2CSK4 and a highly selective inhibitor of PI3K LY294002 were used to clarify the TLR6 signaling pathways. In a conclusion, we discovered that the sensitivity of TLRs in human gastric epithelial cells decreased while after long-time co-culture with H.pylori lysate. Human gastric epithelial cells shows tolerance to H.pylori lysate in TLRs and inflammatory cytokines level. decreased the high level of TLRs and inflammatory cytokines after stimulating GES-1 cells with H.pylori lysate for a short-term. And Tthis process may works through the TLR6/PI3K/NF-κKB axis.

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Coccolithophore viral lysate

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3' isoform specific RNA IP (3' isRIP-seq) for PUF3 RBP in budding yeast

Project description:Exponentially growing cells (OD 0.5-0.8) from the TAP-tagged strain were harvested by ultracentrifugation followed by of washing and flash freezing in liquid nitrogen and breakage in FastPrep24 (ZYmoresearch S6005). The lysate was subjected to IP with Dyna M280 sheep anti rabbit IgG beads for 2h at 4C. An aliquot was taken before the IP as an input control. The IP of the RNA-protein complex was subjected to cleavage by TEV protease to cleave the TAP-tag off the protein. The bound RNA was then precipitated in phenol/chloroform/isoamyl-alcohol and sequenced using 3' T-filling protocol as described by Wilkening et al 2013 to obtain accurate quantification of 3' isoforms. We used the R Bioconductor package DESeq2 (Anders and Huber 2010) to call for differential log change between the Input and the RNA-IP sample and took all those isoforms as differentially regulated which had more than 4-fold change at a FDR of 10% as reported by the package.

2014-11-13 | E-MTAB-2103 | biostudies-arrayexpress

Aspergillus fumigatus EdeA CO-IP

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TMCO1 co-IP elutions and inputs

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AIN-2::GFP IP of mRNAs associated with miRISC in stage sychronized C. elegans populations

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