Canine placenta proteomic analysis

This work aims to map the main protein groups of the native and decellularized (DOI: 10.17632/jkwxmsrn27.1) extracellular matrix of fetal side of canine placentas from 35 days of gestation (end of the middle third of gestation), hypothesizing that the cell removal protocol guarantees a biological matrix capable of maintaining protein groups and structural factors necessary for tissue rebuilding, as well as the processes of cell adhesion and interaction and the restoration of vasculogenesis. For this, non-decellularized (n=3) and SDS-decellularized (n=3) 35-day old fetal part of canine placenta sample were washed, lysed, urea reduced, acetone precipitated, DTT reduced, iodoacetamide alkylated, trypsin digested, and C-18 column purified (accordingly to DOI: 10.1002/9780470559277.ch140272). Finally, 3 µg protein were loaded in OrbitrapFusionLumos spectrometer (Thermo Scientific). Spectra were exported to MaxQuant software (v1.6.10.43) to produce the protein list of each sample, and the LFQ intensity were statistically (p > 0.05) analyzed by Inferno software (v.1.1.6970). After this, proteins related to ECM and cellular junction ontologies were filtered and manually annotated using DAVID Bioinformatics Resources 6.8.