Blood glucose concentrations were measured with a hand-held point-of-care glucose monitor that was previously validated for use in dogs (AlphaTRAK 2; Abbott Animal Health, Abbott Park, IL, USA). Blood samples for cytokine and hormone analysis were collected into chilled EDTA tubes and plasma was separated within 2 h.
Plasma was stored in −20° C and was analyzed in one batch at the end of the study. The analysis was performed in blind fashion. Plasma levels of canine IL6, MCP1, GMCSF, TNFα, insulin, and leptin were measured with MILLIPLEX MAP Canine Cytokine/Chemokine and Canine Endocrine Magnetic Bead Panels, Millipore/Merck, Darmstadt, Germany, Cat No CCYTOMAG-98K). All samples were tested in duplicates.
Paraffin embedded sections of right and left side of injected falciform fat were used for hematoxylin and eosin staining (H&E), as well as immunohistochemistry analysis using rabbit polyclonal antibodies against mouse UCP1 (Abcam, ab10983, react with canine and mouse UCP1) was performed as previously descriebed15 (link). Antibody was used at 1:1000 dilution.
Plasma was stored in −20° C and was analyzed in one batch at the end of the study. The analysis was performed in blind fashion. Plasma levels of canine IL6, MCP1, GMCSF, TNFα, insulin, and leptin were measured with MILLIPLEX MAP Canine Cytokine/Chemokine and Canine Endocrine Magnetic Bead Panels, Millipore/Merck, Darmstadt, Germany, Cat No CCYTOMAG-98K). All samples were tested in duplicates.
Paraffin embedded sections of right and left side of injected falciform fat were used for hematoxylin and eosin staining (H&E), as well as immunohistochemistry analysis using rabbit polyclonal antibodies against mouse UCP1 (Abcam, ab10983, react with canine and mouse UCP1) was performed as previously descriebed15 (link). Antibody was used at 1:1000 dilution.
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