Proteomic analysis of the extracellular and intracellular role of IL-33 in primary human endothelial cells

ABSTRACT: IL-33 is a nuclear cytokine from the IL-1 family that plays important roles in health and disease.Under healthy conditions, IL-33 is constitutively expressed to high levels in the nucleus of producing cells in various human and mouse tissues. The extracellular function of IL-33 cytokine has been well documented, but it remains unclear whether intracellular nuclear IL-33 has additional functions in the nucleus. Here, we used a global proteomic approach based on quantification of 5000 individual proteins by high-resolution mass spectrometry to compare the extracellular and intracellular roles of IL-33 in primary human endothelial cells, a major source of IL-33 protein in human tissues. Large-scale analysis of protein expression was performed either after stimulation of the cells with the IL-33 mature form IL-3395-270 (during 6h or 24h) or after siRNA knockdown of intracellular IL-33 (two experiments, each with a different pool of distinct siRNAs, noted siRNA1 and siRNA2). In each case, proteins were fractionated by 1D SDS-PAGE in 12 gel bands, and label-free quantitative analysis was performed. The present dataset corresponds to the stimulation of human endothelial cells with IL-3395-270 during 6h (IL33_6) or 24h (IL33_24). Cells cultured in the absence of IL-33 were used as control (CTil33). Three independent biological replicates (noted _A, _B, _C) were prepared and analyzed for each condition, leading to 9 different samples. Each of them was fractionated into 12 gel bands analyzed by nanoLC-MS/MS, leading to 108 raw files.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Primary Cell

SUBMITTER: Anne Gonzalez de Peredo  

LAB HEAD: Anne Gonzalez de Peredo

PROVIDER: PXD004117 | Pride | 2016-10-10

REPOSITORIES: Pride