Figure 2

Effect of oxidation agents on HMGB1-induced TNF release. The time dependent inhibitory effect of H₂O₂ exposure on the TNF-stimulating activity of HMGB1 in (A) RAW 267.7 cells and (B) primary human macrophages. (C) The effect of H₂O₂ exposure on the TNF-stimulating activity of LPS in RAW 267.7 cells. Cells were cultured in 96-well plates and were stimulated with HMGB1 or LPS with or without exposure to 50 mmol/L H₂O₂ for 0–120 min. After16 h, TNF released into the cell culture supernatant was measured by ELISA. N = 6–8. *P < 0.05 versus HMGB1 alone without H₂O₂ exposure. (D) The effect of mercury (Hg) on TNF release from RAW 264.7 cells induced by recombinant HMGB1 prepared in the absence of DTT. RAW 264.7 cells were cultured in 96-well plates and exposed to HMGB1 or Hg-HMGB1 over a range of concentrations (0–5 µg/mL). After 16 h, TNF released was measured. N = 3. *P < 0.05 versus HMGB1.