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PSCK9 knockdown in EndoC-bH1 cells time course (DIA secretome time course)

ABSTRACT: EndoC-bH1 cells were incubated with control siRNA or PCSK9 siRNA for 24, 48 and 72 hours. Untreated baseline samples were also included. Unpowered exploratory experiment. Sample size: triplicate per condition.

INSTRUMENT(S): Orbitrap Exploris 480

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Type B Pancreatic Cell, Endocrine Pancreas, Cell Culture

SUBMITTER: Maria Ryaboshapkina

LAB HEAD: Raphael Scharfmann

PROVIDER: PXD027920 | Pride | 2022-04-07

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	PE224.log.txt	Txt
	PE224.params	Other
	PE224.setup.txt	Txt
	PE224.sne	Other
	PE224_2020_GP_PCSK9_1.raw	Raw

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Publications

Characterization of the Secretome, Transcriptome, and Proteome of Human β Cell Line EndoC-βH1.

Ryaboshapkina Maria M Saitoski Kevin K Hamza Ghaith M GM Jarnuczak Andrew F AF Pechberty Séverine S Berthault Claire C Sengupta Kaushik K Underwood Christina Rye CR Andersson Shalini S Scharfmann Raphael R

Molecular & cellular proteomics : MCP 20220402 5

Early diabetes research is hampered by limited availability, variable quality, and instability of human pancreatic islets in culture. Little is known about the human β cell secretome, and recent studies question translatability of rodent β cell secretory profiles. Here, we verify representativeness of EndoC-βH1, one of the most widely used human β cell lines, as a translational human β cell model based on omics and characterize the EndoC-βH1 secretome. We profiled EndoC-βH1 cells using RNA-seq, ...[more]

PMID: 35378291

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PSCK9 knockdown in EndoC-bH1 cells at 72 h (DIA proteomics experiment)

Project description:EndoC-bH1 cells were incubated with control siRNA or PCSK9 siRNA for 72 hours. The experiment was powered based PCSK9 knockdown in a pilot experiment. Sample size: 12x12.

2022-04-07 | PXD027911 | Pride

PSCK9 knockdown in EndoC-bH1 cells at 72 h (DIA secretome experiment)

Project description:EndoC-bH1 cells were incubated with control siRNA or PCSK9 siRNA for 72 hours. The experiment was powered based PCSK9 knockdown in the pilot experiment [PXDxxx (PE083)]. Sample size: 12x12.

2022-04-07 | PXD027913 | Pride

PSCK9 knockdown in EndoC-bH1 cells at 72 h (pilot TMT experiment)

Project description:EndoC-bH1 cells were incubated with control siRNA or PCSK9 siRNA for 72 hours. Unpowered exploratory experiment. Sample size: 3x3.

2022-04-07 | PXD027921 | Pride

Effect of control siRNA knockdown on EndoC-bH1 cell lysate proteome (pilot TMT experiment)

Project description:EndoC-bH1 cells proteomics was assessed in untreated baseline condition and after treatment with control siRNA at 24, 48 and 72 hours. Unpowered exploratory experiment. Sample size: triplicate per condition.

2022-04-07 | PXD027898 | Pride

Mass spectrometry analysis of mouse hematopoietic stem cells and their progenitors reveals differential expression within and between proteome and transcriptome throughout adult and aged hematopoiesis

Project description:mass spectrometry analysis of aged and adult hematopoietic stem cells, multipotent progenitors and oligopotent progenitors

2021-07-24 | PXD017442 | Pride

PCSK9 knockdown in EndoC-bH1 cells at 72 hours

Project description:The aim of the study was to characterize the role of PCSK9 in human beta cells. We performed siRNA-mediated knockdown of PCSK9 in human beta cell line EndoC-bH1 and compared the expression profiles against control siRNA-treated cells.

2022-03-31 | GSE182016 | GEO

Spatiotemporal profiling of the bovine oviduct fluid proteome around the time of ovulation

Project description:Unraveling the composition and regulation of the oviduct fluid (OF) is crucial to understand the microenvironment in which sperm capacitation, fertilization and early embryo development take place. Therefore, our aim was to determine the spatiotemporal changes in the OF proteome according to the anatomical region of the oviduct (ampulla vs. isthmus), the proximity of the ovulating ovary (ipsilateral vs. contralateral side) and the peri-ovulatory stage (pre-ovulatory or Pre-ov vs. post-ovulatory or Post-ov). Oviducts from adult cyclic cows were collected at a local slaughterhouse and pools of OF were analyzed by nanoLC-MS/MS and label-free protein quantification (n=32 OF pools for all region stage side conditions). A total of 3,760 proteins were identified in the OF, of which 37% were predicted to be secreted. The oviduct region was the major source of variation in protein abundance, followed by the proximity of the ovulating ovary and finally the peri-ovulatory stage. Differentially abundant proteins between regions, stages and sides were involved in a broad variety of biological functions, including protein binding, response to stress, cell-to-cell adhesion, calcium homeostasis and the immune system. This work highlights the dynamic regulation of oviduct secretions and provides new protein candidates for interactions between the maternal environment, the gametes and the early embryo.

2022-03-03 | PXD030915 | Pride

Carnitine O-octanoyltransferase is a novel inducer of vascular calcification via promoting fatty acid metabolism and mitochondrial dysfunction

Project description:Objective – Vascular calcification is a critical pathology associated with increased cardiovascular event risk, but there are no FDA-approved anti-calcific therapies. We hypothesized and validated that an unbiased screening approach would identify novel mediators of human vascular calcification. Approach and Results – We performed an unbiased quantitative proteomics and pathway network analysis that identified increased carnitine O-octanoyltransferase (CROT) in calcifying primary human coronary artery smooth muscle cells (SMCs). Additionally, human carotid artery atherosclerotic plaques contained increased immunoreactive CROT near calcified regions. CROT siRNA reduced fibrocalcific response in calcifying SMCs. In agreement, histidine 327 to alanine point mutation inactivated human CROT fatty acid metabolism enzymatic activity and suppressed SMC calcification. CROT siRNA restored mitochondrial proteome alterations and suppressed mitochondrial fragmentation in calcifying SMCs. Lipidomics analysis of SMCs incubated with CROT siRNA revealed increased eicosapentaenoic acid, a vascular calcification inhibitor. CRISPR/Cas9-mediated Crot deficiency in low-density lipoprotein receptor-deficient mice reduced aortic and carotid artery calcification without altering bone density, or liver and plasma cholesterol and triglyceride concentrations. Conclusions – CROT is a novel inducer of vascular calcification via promoting fatty acid metabolism and mitochondrial dysfunction, as such CROT inhibition has strong potential as an anti-fibrocalcific therapy.

2021-02-26 | PXD022575 | Pride

Mitochondrial proteome analysis during MceC expression time-course

Project description:Mitochondria were isolated from tetracycline-inducible control (EV) HEK293 cells or HEK293 cells expressing the C. burnetii effector protein MceC for 1, 4 and 8 hours.

2020-11-12 | PXD019337 | Pride

Quantitative mitochondrial proteome analysis during MceC expression time-course

Project description:Mitochondria were isolated from tetracycline-inducible control (EV) HEK293 cells or HEK293 cells expressing the C. burnetii effector protein MceC for 1, 4 and 8 hours.

2020-11-12 | PXD021704 | Pride

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