2.6. Determination of Pro-Inflammatory Cytokines Production

MCP-1, IL-6, IL-1β, and TNF-α levels produced by adipocytes and SVCs were determined using Mouse ELISA MCP-1, IL-6, IL-1β, and TNF-α kits (BD Bioscience, San Diego, CA, USA), following the manufactures’ instructions. The absorbance was measured at 450 nm with a microplate spectrophotometer (Spectramax190, Molecular devices, CA, USA).

To normalize pro-inflammatory cytokine levels for the number of adipocytes, the total DNA from adipocytes was measured using a Quick-DNA™ Miniprep Plus kit (Zymo research, Irvine, CA, USA).