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. 2016 May 19;1(7):e85375. doi: 10.1172/jci.insight.85375

Figure 3. Haptoglobin attenuates HMGB1-induced TNF expression and secretion in macrophages.

Figure 3

(A) Upper: RAW 264.7 cells in 96-well culture plates were stimulated with HMGB1 (2 μg/ml) plus various amounts of haptoglobin as indicated for 16 hours. TNF release was measured (n = 5 experiments). Lower: For TNF mRNA measurement, RAW 264.7 cells were incubated with HMGB1 and various amounts of haptoglobin for 12 hours. TNF mRNA expression in the cells was measured by real-time quantitative PCR. Housekeeping gene HPRT was used as a control for equal loading and integrity of RNA (n = 8 experiments). *P < 0.05, ***P < 0.001 vs. HMGB1 alone (t test). (B) Haptoglobin inhibits HMGB1-induced secretion of IL-8 and TNF in human macrophages. Primary human macrophages were incubated with HMGB1 (2 μg/ml) in the presence of haptoglobin at various concentrations as indicated for 16 hours. TNF and IL-8 released were measured. Data are from 8 (for TNF) or 5 (for IL-8) experiments. **P < 0.01, ***P < 0.001 vs. HMGB1 alone (t test).