Project description:T cell receptor (TCR) signaling is essential for the function of T cells. Here we monitor and quantify the dynamics of phosphorylation sites in primary CD4+ T cells during the first 10 minutes after antibody-based TCR stimulation. To capture the earliest molecular events induced by TCR engagement we prepared unstimulated and stimulated T cells (15, 30, 120, 300 or 600 seconds after stimulation). Cells were then lysed in urea, proteins were digested into tryptic peptides, and phosphorylated peptides were enriched using titanium (TiO2) beads. In most experiments a further enrichment step was performed using phospho-Tyrosine (pTyr) immuno-precipitation. We performed 4 independent experiments, each containing 6 time-points of stimulation. This dataset contains the raw LC-MS/MS files of the pre-enriched peptide samples (SP, 24 samples) and the matching TiO2-enriched peptides (TiO2, 24 samples), as well as the eluates from further pTyr immuno-precipitation from 3 experiments (Ptyr, replicates 1, 3, 4: 18 samples). All samples were analyzed in triplicate or quadruplicate nanoLC-MS runs.
Project description:HUVECs were stimulated and samples were prepared after 0 and 30 min. Chromatin interaction mediated by active RNA polymerase II was detected by ChIA-PET.
Project description:Subcellular proteomics analysis of human T-cells in response to T-cell receptor stimulation. High Resolution Isoelectric Focusing (HiRIEF) LC-MS/MS and relative quantification by TMT 10-plex was used to analyze subcellular fractions (Cytosol, Nuclear and Mitochondria-Small Organelle-Membrane) of Human T-cells from 3 separate donors stimulated with T-cell receptor stimulation for 0, 15 minutes or 60 minutes. The data was obtained from 3 separate TMT10 plex sets which included: 126 – Cytosol, unstimulated; 127N – Cyto, 15 min; 127C – Cyto, 60 min; 128N – Mitochondrion-Small Organelle-Membrane, unstimulated; 128C – Mito-SmO-Mem, 15 min; 129N – Mito-SmO-Mem, 60 min; 129C – Nuclear, unstimulated; 130N – Nuc, 15 min; 130C– Nuc, 60 min; 131 – pooled internal control of all samples.
Project description:The series was designed to obtain gene expression patterns of TNF-alpha stimulated CaCo-2 cells. Keywords: TNF-alpha stimulation This series represents a group of 18 samples in 2 groups: 10 samples derived from not stimulated CaCo-2 cells, 8 samples from CaCo-2 cells stimulated with TNF-alpha in final concentration of 5 ng/ml for 60 min.
Project description:HUVECs were stimulated and samples were prepared after 0 and 30 min. Chromatin interaction mediated by active RNA polymerase II was detected by ChIA-PET. Samples were made afrer 0 and 30 min after TNF alpha stimulation.
Project description:Interventions: experimental group:Patients in the experimental group were stimulated with transcranial direct current stimulation using anodal stimulation of the left dorsolateral prefrontal lobe and cathodal stimulation of the right dorsolateral prefrontal lobe, with a stimulation duration of 20 min and a stimulation current of 2 mA.;control group:In the control group, the stimulating electrodes were placed on the corresponding site, and the instrument was turned on to give a short climbing pseudo-stimulation for 30 s. After that, the instrument was turned off.
Primary outcome(s): Incidence of perioperative anxiety
Study Design: Parallel
Project description:T-cell receptor (TCR) signaling is essential for the function of T cells and negatively regulated by the E3 ubiquitin-protein ligases CBL and CBLB. Here we combine mouse genetics and affinity purification coupled to quantitative mass spectrometry to monitor the dynamics of the CBL and CBLB signaling complexes that assemble in normal T cells over 600 seconds of TCR stimulation. We identify many previously known CBL and CBLB interacting partners, as well as a majority of proteins that have not yet been implicated in those signaling complexes. We exploit correlations in protein association with CBL and CBLB as a function of time of TCR stimulation for predicting the occurrence of direct physical association between them. The dataset is divided in two distinct subsets corresponding to the Cbl and to the Cblb interactomes. Each of them contains mass spectrometry results from the analysis of 10 different conditions of AP-MS purifications (based on affinity purification on Streptactin beads of One-Strep-tagged proteins) starting from CD4+ T cells which were either non stimulated or stimulated with anti-CD3 and anti-CD4 antibodies as follows: CBL interactome: - CBL-OST transgenic mice, CD4+ T cells non stimulated (noted Cbl_0) - CBL-OST transgenic mice, CD4+ T cells stimulated 30s (noted Cbl_30) - CBL-OST transgenic mice, CD4+ T cells stimulated 120s (noted Cbl_120) - CBL-OST transgenic mice, CD4+ T cells stimulated 300s (noted Cbl_300) - CBL-OST transgenic mice, CD4+ T cells stimulated 600s (noted Cbl_600) - WT mice, CD4+ T cells non stimulated (noted WT_0) - WT mice, CD4+ T cells stimulated 30s (noted WT_30) - WT mice, CD4+ T cells stimulated 120s (noted WT_120) - WT mice, CD4+ T cells stimulated 300s (noted WT_300) - WT mice, CD4+ T cells stimulated 600s (noted WT_600) For the CBL interactome, 3 biological replicates were prepared for these 10 different conditions (noted Ech1, Ech2, Ech3), yielding 30 analyzed samples. Three technical nanoLC-MS runs were acquired for each sample (noted R1, R2, R3), leading to 90 nanoLC-MS raw files. CBL interactome: - CBLB-OST transgenic mice, CD4+ T cells non stimulated (noted Cblb_0) - CBLB-OST transgenic mice, CD4+ T cells stimulated 30s (noted Cblb_30) - CBLB-OST transgenic mice, CD4+ T cells stimulated 120s (noted Cblb_120) - CBLB-OST transgenic mice, CD4+ T cells stimulated 300s (noted Cblb_300) - CBL-OST transgenic mice, CD4+ T cells stimulated 600s (noted Cblb_600) - WT mice, CD4+ T cells non stimulated (noted WT_0) - WT mice, CD4+ T cells stimulated 30s (noted WT_30) - WT mice, CD4+ T cells stimulated 120s (noted WT_120) - WT mice, CD4+ T cells stimulated 300s (noted WT_300) - WT mice, CD4+ T cells stimulated 600s (noted WT_600) For the CBLB interactome, 4 biological replicates were prepared for these 10 different conditions (noted S1, S2, S3, S4), yielding 40 analyzed samples. For series S1 to S3, 2 technical nanoLC-MS runs were acquired for each sample (noted R1, R2) and for serie S4, 3 technical nanoLC-MS runs were acquired for each sample (noted R1, R2, R3), leading in total to 90 nanoLC-MS raw files.
Project description:The series was designed to obtain gene expression patterns of TNF-alpha stimulated THP-1 cells. Keywords: TNF-alpha stimulation This series represents a group of 15 samples in two groups: 7 samples derived from not stimulated THP-1 cells, 8 from THP-1 cells stimulated with TNF-alpha in a final concentration of 5 ng/ml for 60 min.
Project description:Blood samples were collected from all participants in 10-mL EDTA-coated Vacutainer tubes. Plasma was separated by centrifugation at 3,000 rpm for 10 min at room temperature (25°C) within 2 h after blood collection and then centrifuged at 13,000 rpm for 10 min at 4°C to remove debris. Isolated plasma samples were stored at -80°C until use. EV RNA were isolated by affinity-based binding to spin columns using an exoRNeasy Serum/Plasma kit (Qiagen, Hilden, Germany). RNA libraries were prepared for sequencing using SMARTer® Stranded Total RNA-Seq Kit - Pico Input Mammalian. RNA-seq was performed by the Illumina sequencing platform on a 150-bp paired-end run.
Project description:C2C12 myotubes at day 7 of differentiation were stimulated with a single session of SIT (6 x 30s with 4 min at rest) and were immediately after the stimulation treated or not with 10 uM S107 drug for 72h. The myotubes were then harvested at 72h post stimulation (for SIT condition) and 72h post stimulation and S107 treatment (for SIT S107 condition) for mass spectrometry analysis using the proteomics approach. N = 5 independent myotube wells per condition.