Figure 5

LPS induced TNFalpha protein secretion requires co-factors and is dependant upon the CD14/TLR4 pathway. Panel A: FBS effect on LPS induced TNFalpha secretion. The concentrations of TNFalpha (pg/mL) in the medium of mature adipocyte cultures treated or not with increasing concentrations of FBS and 1 μg/mL of LPS, were determined at 6 hours. The graph represents the mean ± SE of the results from two patients. ***P < 0.001%, *P < 0.05%, versus control cells. Panel B: Effect of TLR4 blocking antibody on TNFalpha secretion. The concentrations of TNFalpha (pg/mL) in the medium of mature adipocyte cultures treated with 1 μg/mL of LPS, with or without 1 h of pre-incubation with a blocking anti-TLR4 antibody (20 μg/mL, Clinisciences, Montrouge, France, clone HTA125) were determined at 6 hours. The graph represents the mean ± SE of the results from two patients. ***P < 0.001%, versus cells treated with LPS alone. Panel C: Effect of CD14 blocking antibody on TNFα secretion. The concentrations of TNFα (pg/mL) in the medium of mature adipocyte cultures treated with 1 μg/mL of LPS, with or without 1 h of pre-incubation with a blocking anti-CD14 antibody (5 μg/mL, Hycult biotechnology, clone 18D11) were determined at 6 hours. The graph represents the mean ± SE of the results from one patient (n = 6 for each condition), representative of two experiments on two different patients. ***P < 0.001%, versus cells treated with LPS alone.