Fig. 3. TLR2/4 gene expression is enhanced by the synergy between oxidative stress and obesity-associated factors. PBMC were incubated with leptin, LPS, oleate, or palmitate, with or without oxidative stress (H₂O₂). Total RNA was extracted, and TLR2/4 gene expression was assessed by real-time RT-PCR as described in materials and methods. The data (mean±SEM) obtained from 5 independent determinations with similar results show significant upregulation of: (A) TLR2 expression in cells treated with leptin+H₂O₂ (41.53±1.34 fold), LPS+H₂O₂ (39.50±3.75 fold), oleate+H₂O₂ (40.27±1.18 fold), or palmitate+H₂O₂ (45.10±1.07 fold); and (B) TLR4 expression in cells treated with leptin+H₂O₂ (19.50±1.04 fold), LPS+H₂O₂ (18.40±0.64 fold), oleate+H₂O₂ (17.78±0.06 fold), or palmitate+H₂O₂ (21.90±1.10 fold) compared to respective controls treated with H₂O₂ alone.