Proteomic survey of Oxytricha trifallax macronuclei and micronuclei
Ontology highlight
ABSTRACT:
INSTRUMENT(S): Orbitrap Fusion
ORGANISM(S): Oxytricha Trifallax (ncbitaxon:1172189)
SUBMITTER: 
Laura Landweber
PROVIDER: MSV000090958 | MassIVE | Thu Dec 22 13:58:00 GMT 2022
REPOSITORIES: MassIVE
Similar Datasets
Project description:Part I - SI-NET tumors High Resolution Isoelectric Focusing (HiRIEF) LC-MS and relative quantification by iTRAQ 8-plex was used to analyze 14 small intestinal neuroendocrine tumors (SI-NETs). The data was obtained from two separate TMT10plex sets and linked by a single internal pooled standard. The internal pooled standard was made by combining equal aliquots of the tryptic peptide mixtures from each of the 14 tissue samples. iTRAQ set1 was composed of 7 SI-NET samples, all from different individuals, and internal pooled standard labelled as follows: Channel 113 (sample Screen-1 with liver metastasis), Channel 114 (sample Screen-8 with liver metastasis), Channel 115 (sample Screen-3 with liver metastasis), Channel 116 (sample Screen-2 with liver metastasis), Channel 117 (sample Screen-5 no liver metastasis), Channel 118 (sample Screen-4 no liver metastasis), Channel 119 (sample Screen-10 no liver metastasis), Channel 121 (internal pooled standard). iTRAQ set2 was composed of 7 SI-NET samples, all from different individuals, and internal pooled standard labelled as follows: Channel 113 (sample Screen-7 with liver metastasis), Channel 114 (sample Screen-11 with liver metastasis), Channel 115 (sample Screen-13 with liver metastasis), Channel 116 (sample Screen-6 no liver metastasis), Channel 117 (sample Screen-12 no liver metastasis), Channel 118 (sample Screen-9 no liver metastasis), Channel 119 (sample Screen-14 no liver metastasis), Channel 121 (internal pooled standard). Part II - time course profiling in cell lines High Resolution Isoelectric Focusing (HiRIEF) LC-MS and relative quantification by TMT 10-plex was used to analyze cellular response to the neddylation inhibitor pevonedistat (MLN4924) at different timepoints in two SI-NET (small intestinal neuroendocrine tumors) cell lines. The data was obtained from two separate TMT 10-plex experiments. TMT set1 includes a time course experiment upon pevonedistat treatment of CNDT2 cells with harvests at 2h, 6h, 12h and 24h after treatment as well as of untreated control cells. Isobaric tag labelling of peptide samples with TMT10plex was used as follows. Biological duplicate controls (TMT channels 126, 127N), duplicate 2h (127C, 128N), duplicate 6h (128C, 129N), duplicate 12h (129C, 130N) and duplicate 24h (130C, 131) samples were employed. TMT set2 includes a time course experiment upon pevonedistat treatment of HC45 cells with harvests at 2h, 6h, 12h and 24h after treatment as well as of untreated control cells. Isobaric tag labelling of peptide samples with TMT10plex was used as follows. Biological duplicate controls (TMT channels 126, 127N), duplicate 2h (127C, 128N), duplicate 6h (128C, 129N), duplicate 12h (129C, 130N) and duplicate 24h (130C, 131) samples were employed.
2020-12-09 | PXD009877 | Pride
Project description:Gastric ECL cells and parietal (PC) cells in normal conditions and gastric ECL cells after 24 hrs fasting were purified using a combination of counter-flow elutriation, nycodenz gradient and FACS based on acridine orange labeling of histamine containing vesicles (ECL cells) or autofluorescense (PC cells) to homogeneity. cRNA probes from this purified cell suspensions and from mixed gastric homogenates were hybridized to whole rat genome expression oligonucleotide microarrays. We imported each channel separately (normalized intensity of Cy3 labeled samples = gProcessedSignal and of Cy5 labeled samples = rProcessedSignal) from the original Agilent Feature Extraction data files (submitted as raw data) after slide scanning from all slides into a Genespring 7.3 microarray database warehouse. Genespring’s built in statistical module was used to compare all channels to each other. The normalization used was against specific samples (the 5 channels of Gastric mucosal scrapings). Genespring identifies genes differentially expressed in the ECL cell samples and the parietal samples and also calculates the significance of difference (p-value). Keywords: rat whole genome expression microarray analysis (transcriptome)
2006-11-22 | GSE6204 | GEO
Project description:We used mass spectrometry-based proteomics to perform a phosphoproteomics analysis of selinexor-treated ex vivo AML patient samples (n=20) and 4 AML cell lines (PL-21, NOMO-1, GDM-1 and MV4-11). For quantitative phosphoproteomics, we used a tandem mass tag (TMT)-based approach. Conditions for the TMT 11-plex (Ex vivo samples) and TMT 8-plex (AML cell lines) setups are specified below. Cells were treated with 1 M selinexor (also known as KPT-330) or 0.1% DMSO for 6 hours. The experimental treatment conditions and TMT11-plex (ex vivo samples) and TMT8-plex (cell lines) labeling schemes are specified below: TMT-11-plex TMT channel Patient no Treatment Set 1 126 Ex_24 DMSO Set 1 127N Ex_24 Selinexor Set 1 127C Ex_25 DMSO Set 1 128N Ex_25 Selinexor Set 1 128C Ex_34 DMSO Set 1 129N Ex_34 Selinexor Set 1 129C Ex_41 DMSO Set 1 130N Ex_41 Selinexor Set 1 130C Ex_44 DMSO Set 1 131N Ex_44 Selinexor Set 1 131C Mix pool Mix pool Set 2 126 Ex_9 DMSO Set 2 127N Ex_9 Selinexor Set 2 127C Ex_18 DMSO Set 2 128N Ex_18 Selinexor Set 2 128C Ex_27 DMSO Set 2 129N Ex_27 Selinexor Set 2 129C Ex_29 DMSO Set 2 130N Ex_29 Selinexor Set 2 130C Ex_36 DMSO Set 2 131N Ex_36 Selinexor Set 2 131C Mix pool Mix pool Set 3 126 Ex_8 DMSO Set 3 127N Ex_8 Selinexor Set 3 127C Ex_13 DMSO Set 3 128N Ex_13 Selinexor Set 3 128C Ex_14 DMSO Set 3 129N Ex_14 Selinexor Set 3 129C Ex_19 DMSO Set 3 130N Ex_19 Selinexor Set 3 130C Ex_40 DMSO Set 3 131N Ex_40 Selinexor Set 3 131C Mix pool Mix pool Set 4 126 Ex_6 DMSO Set 4 127N Ex_6 Selinexor Set 4 127C Ex_15 DMSO Set 4 128N Ex_15 Selinexor Set 4 128C Ex_28 DMSO Set 4 129N Ex_28 Selinexor Set 4 129C Ex_39 DMSO Set 4 130N Ex_39 Selinexor Set 4 130C Ex_42 DMSO Set 4 131N Ex_42 Selinexor Set 4 131C Mix pool Mix pool AML cell lines: TMT-8-plex TMT channel Treatment Replicate GDM-1 126 DMSO 1 GDM-1 127N DMSO 2 GDM-1 127C DMSO 3 GDM-1 128N DMSO 4 GDM-1 128C Selinexor 1 GDM-1 129N Selinexor 2 GDM-1 129C Selinexor 3 GDM-1 130N Selinexor 4 MV4-11 126 DMSO 1 MV4-11 127N DMSO 2 MV4-11 127C DMSO 3 MV4-11 128N DMSO 4 MV4-11 128C Selinexor 1 MV4-11 129N Selinexor 2 MV4-11 129C Selinexor 3 MV4-11 130N Selinexor 4 NOMO-1 126 DMSO 1 NOMO-1 127N DMSO 2 NOMO-1 127C DMSO 3 NOMO-1 128N DMSO 4 NOMO-1 128C Selinexor 1 NOMO-1 129N Selinexor 2 NOMO-1 129C Selinexor 3 NOMO-1 130N Selinexor 4 PL-21 126 DMSO 1 PL-21 127N DMSO 2 PL-21 127C DMSO 3 PL-21 128N DMSO 4 PL-21 128C Selinexor 1 PL-21 129N Selinexor 2 PL-21 129C Selinexor 3 PL-21 130N Selinexor 4
2022-08-07 | PXD017660 | Pride
Project description:The goal of this study was to identify ion channels, specifically transient receptor potential cation channel A (trpA1) channels, that were highly expressed and enriched in nociceptive sensory neurons of Drosophila larvae. In class IV da sensory neurons, we find that TrpA1 is the most highly expressed trpA1 channel of the 14 trpA1 channels in Drosophila, and that its expression is highly enriched when compared to the whole animal transcriptome.
2017-11-06 | GSE99711 | GEO
Project description:5 rats were offered food containing 40mM Li/Kg dry weight for 4 weeks, and 5 control rats obtained standard food. The RNA from the inner medulla of the Li-treated rats was labeled red (channel 1), and from the control rats labeled green (channel 2). The samples from Li-treated rats 1+2 and control rats 1+2 were hybridized to array Li-NDI 1. The samples from Li-treated rats 3+4 and control rats 3+4 were hybridized to array Li-NDI 2. The samples from Li-treated rat 5 and control rat 5 were hybridized to array Li-NDI 3. Keywords: parallel sample
2004-09-01 | GSE1341 | GEO
Project description:Microarray Expression profiles from 58 samples comprising of 3 normals and 55 gastric cancers is used for molecular subclassification of Gastric Cancers. Data Generation and Processing: The ratiometric data here is given by Ch1(Cy3)/Ch2(Cy5) where Ch2 is Universal Reference RNA from Stratagene. Two different chips were used (13K and 18k); The ratio metric is given by (Biological sample / Universal Reference). The Biological Samples are labeled with Cy3 (Green) and the Universal Reference RNA is always labeled with Cy5 (Red). Two types of spots were excluded a) Spots for which Foreground intensity was lower from background by lesser than 10 fluoresence units. b) Spots that couldnt be identified reliably by the scanner (Arrayworx) software within the "region of interest" masked by the user. The 2 channels for each array are equalized by multiplying the reference channel Ch2(Cy5) by a constant which is derived by (Total Cy3 / Total Cy5). Subsequently, all expression ratios from each array is normalized to have median expression value = 1. Keywords: other
2005-07-25 | GSE2637 | GEO
Project description:Pannexin 1 (Panx1) channels can be activated by alpha 1 adrenoceptor-induced pathways for the sympathetic regulation of blood pressure; however, the molecular mechanisms for this form of Panx1 channel activation remain elusive. To identify potential acetyl-lysine residue(s) of Panx1 channels that might be involved in this receptor-mediated Panx1 channel activation, we performed mass-spectrometry on Strep-tagged Panx1 proteins precipitated from the whole cell lysate of HEK293T cells after stable isotope labeling by amino acids in cell culture (SILAC). Those HEK293T cells were transiently transfected with alpha1D adrenoceptors and human Panx1-Strep, with or without phenylephrine stimulation. Equal amounts of light-labeled (control) and heavy-labeled (phenylephrine stimulated) cell lysates were pooled, and Panx1 proteins were precipitated by Strep-Tactin beads, followed by LC-MS-MS analysis. From three biological replicates, we identified a consistent, albeit modest, reduction of acetylation level at K140, following phenylephrine stimulation. Whereas the acetylation level of other lysine residues (K321, K374, K381, K409) were variable among three replicates or were unaffected by phenylephrine treatment. These results implicate Panx1 K140 as a potential regulatory site for receptor-mediated channel activation via an acetylation-deacetylation mechanism.
2021-07-16 | PXD025912 | Pride
Project description:Using RNA sequencing of FACS sorted retro-labeled sensory neurons innervating tongue tissue, we determined changes in transcriptomic profiles in males and female mice under naïve as well as tongue-tumor bearing conditions Our data revealed the following interesting findings: 1) Naïve female neurons innervating the tongue exclusively expressed immune cell markers such as Csf1R, C1qa and others, that weren’t expressed in males. 2) Male neurons were more tightly regulated than female neurons upon tumor growth; 3) While very few differentially expressed genes (DEGs) overlapped between males and females post-tumor growth, several biological processes (BPs) were similar between two sexes. However, additional distinct processes were sex-specific; 4) Post-tumor growth, male DEGs contained an equal mix of transcription factors, ligands, growth factors, receptors and channels, whereas female DEGs predominantly contained channels/receptors, enzymes, cytokines and chemokines.
2023-08-15 | GSE220448 | GEO
Project description:Investigating the impacts on MS data quantitation reproducibility when loading differential amounts of peptides, ranging from 20 to 400 ug, across channels of TMT 11 multiplexes. PTRC_Exp9 files represent global proteome and phosphoproteome data generated from PBMCs isolated from AML patients, acquired through 12 fractions and 6 fractions per plex, respectively. In this experiment, TMT channels were loaded with varying peptide quantities ranging from 20 to 400 ug. PTRC_Exp12 files represent proteomic and phosphoproteomic data from the MOLM-14 AML cell line, also acquired across 12 global proteomics fractions and 6 phosphoproteomics fractions per plex. In this experiment, all TMT channels were loaded with equivalent quantity of peptides (400 ug). Samples were reduced with dithiothreitol, alkylated with iodoacetamide, and double-digested with Lys-C and trypsin. Digested peptides were desalted with C18 SPE columns, labeled with TMT11 isobaric tags, and fractionated by high pH reverse phase separation. After aliquots were removed for global proteomics (12 fractions per plex), samples were concatenated and IMAC was used to enrich phosphopeptides (6 fractions per plex). LC-MS/MS measurements were acquired on an Orbitrap Fusion Lumos mass spectrometer.
2020-11-05 | MSV000086417 | MassIVE
Project description:To explore the molecular basis of the distinct intrinsic membrane properties and other dstinguishing features of functionally defined DRG neuron subtypes, we bulk-sequenced RNA at high depth of genetically-labeled DRG neurons to generate transcriptome profiles of eight major DRG neuron subtypes. The trancriptome profiles revealed differentially expressed and functionally relevant genes, including voltage-gated ion channels. Guided by the transcriptome pofiles, electrophysiological analyses using pharmacological and genetic manipulations as well as computational modeling of DRG neuron subtypes were undertaken to assess the functions of select voltage-gated potassium channels (Kv1, Kv2, Kv3, and Kv4) in shaping action potential (AP) waveforms and firing patterns of the DRG neuron subtypes. Our findings show that the transcriptome profiles have predictive value for defining ion channel contributions to sensory neuron subtype-specific intrinsic physiological properties.
2019-05-16 | GSE131230 | GEO