Proteomics raw data of serum extracellular vesicles from bitches with mammary neoplasia
Description
These data are related to research investigating proteomics in serum extracellular vesicles (EVs) from bitches with mammary neoplasia for comparison to young bitches with no history of neoplasia. Our hypothesis was that neoplasia groups have a different abundance of proteins from the control bitches. We collected blood, separated the serum, isolated the EVs, and then analysis from the EVs' protein cargo. Six proteins had differences in abundance among the groups.
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Serum samples were collected and separated into 3 groups: Control (young females without a history of neoplasms, n = 10), 2 groups composed of bitches with mammary neoplasia, group I (lower malignancy neoplasia, n = 13), and group II (higher malignancy neoplasia, n = 7) classified according to histological characteristics (Cassali et al. 25,26 and Goldschmidt et al. 27). Serum was prepared for a series of 4 centrifugations. The first was used to separate the blood serum (~720xg for 10 minutes at RT), followed by 3x centrifugations at 4 °C (300xg/10 min, 2,000xg/10 min, and 16,500xg/30 min) (Théry et al. 29). Thus, EVs were isolated by size exclusion chromatography, quantified, and characterized by concentration and size distribution (nanoparticle tracking analysis), and size (MET). The proteins were extracted using a RIPA buffer and sonication, and the samples were submitted to tryptic digestion (Shevchenko et al. 35). Mass spectrometry was conducted in an ESI-Q-TOF MS/MS shotgun approach. Spectra were acquired (MassLynx™ v.4.1 software) and analyzed (Mascot Distiller 2.4.0.0 software) to determine by the exponentially modified protein abundance index (emPAI). The results were validated in the Protein Pilot 4.0 software using the taxonomy Canis lupus familiaris (UP000002254, UniProtKB). Comparison of the size and concentration of EVs was performed by one-way ANOVA/Tukey test and Kruskal-Wallis/ Tukey test, respectively (P < 0.05). The data were normalized for proteomics to exclude outliers and analyzed by a non-hierarchical clustering (MetaboAnalyst 5.0 software). one way ANOVA/Fisher's LSD test was also used to find the different abundances of proteins among groups (FDR < 0.05).
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Funding
The São Paulo Research Foundation (FAPESP)
2021/07318-2
Coordination for the Improvement of Higher Education Personnel (CAPES)
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