Inhibition of Inflammatory Cytokines in Adipocytes

When confluence was reached, preadipocytes (n = 6 wells per group) were cultured for 24 h in preadipocyte growth medium, which was replaced with 25% MacCM (diluted in RPMI-1640) to elicit inflammatory responses, together with IL-6 antibody (300, 350 and 450 ng/ml) or IL-1β antibody (15 μg/ml as previously established [18 (link)]) (R&D Systems, UK) to block IL-6 or IL-1β action by neutralization for a further 24 h before cell and supernatant collection. Isotype control mouse IgG at the same concentrations confirmed that non-specific binding did not block inflammatory responses (data not shown). The control received a mock treatment of 25% preadipocyte growth medium (diluted in RPMI-1640).