Identification of proteins with increased expression in adipocyte CM in the presence of ECM (AE-CM) revealed a top 4 list. (A) SDS-PAGE of adipocyte CM (A-CM) and AE-CM reveals two protein bands exclusive to AE-CM. Molecular weight markers are shown to the left of the gel and the two differentially expressed bands are indicated with arrows. Data are representative of two independent replicates; (B) in a semi-quantitative proteomic analysis, 141 unique proteins were found in A-CM, 48 distinctive proteins were exclusively found in AE-CM, and 219 proteins were found in both conditions; (C) cellular localization of differentially expressed proteins. Note that only 19 are shown as we specifically examined secretory vesicle, extracellular space and ECM; (D) relative abundance of 4 differentially secreted proteins by number of peptide spectrum matches. Two independent replicates were analyzed. +/–: cultures without (–) or with (+) the Matrigel-based ECM
We would like to thank Andrada Tarta (Department of Biomedical Sciences, University of Guelph), for her valuable contribution to the optimization of the adipocyte differentiation protocol at the A.V.P lab.
Author contributions
CMVS: Conceptualization, Investigation, Formal analysis, Validation, Visualization, Writing—original draft, Writing—review & editing. GK: Conceptualization, Investigation, Formal analysis, Writing—review & editing. NKP, EE and AVVC: Investigation, Formal analysis. SLC and AMVP: Conceptualization, Methodology, Writing—review & editing, Supervision, Project administration, Funding acquisition. All authors read and approved the submitted version.
Conflicts of interest
The authors declare that they have no conflicts of interest.
This work was funded by a Cancer Research Society operating grant to SC and AVP [22130]; an Internal Faculty of Science, Memorial University of Newfoundland award to SC; a Natural Sciences and Engineering Research Council (NSERC) of Canada grant to AVP [#401248]; CMVS was supported by an Art Rouse Memorial Master of Science scholarship from the Ontario Veterinary College (OVC); GK was supported by a Carol Ann Cole Comfort Heart Summer Studentship via the Beatrice Hunter Cancer Research Institute and is supported by a CGS-M award from the Canadian Institutes of Health Research (CIHR). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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