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. Author manuscript; available in PMC: 2013 Aug 15.
Published in final edited form as: J Immunol. 2012 Jul 6;189(4):2017–2022. doi: 10.4049/jimmunol.1103623

Figure 3. Involvement of induction of ROS and activation of Syk in the mechanisms of synergy.

Figure 3

BMDMs from C57BL/6 mice were washed 3 times and pre-treated with (A) Serum free media (SFM), or NAC (10mM) or DPI (10 μM) or allopurinol (10 μM) or L-NAME (100 μM) or (B) piceacetannol at indicated concentrations for 30 min. Cells were then further treated overnight with serum-free medium or with 4 μg/ml of HMGB1 in the absence or presence of 30 μg/ml mouse hemoglobin (Hb). Concentrations of TNF in the supernatants were determined by ELISA. The results denote the mean ± s.e and are representative of more than four independent experiments. * P < .05, ** P < .01, compared between cells treated with and without hemoglobin in the presence of serum free media or the same type of inhibitor. In panel A, † P < .001, compared between cells treated with HMGB1 and hemoglobin in the absence and presence of DPI. In panel B, P < .001 for the dose effect of piceacetannol on cells treated with HMGB1 and hemoglobin.