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. Author manuscript; available in PMC: 2012 Aug 27.
Published in final edited form as: J Immunol. 2010 Aug 27;185(7):4385–4392. doi: 10.4049/jimmunol.1000803

Figure 4.

Figure 4

HMGB1 release from Salmonella-infected macrophages requires bacterial flagellin, a functional type III secretion and the Ipaf inflammasome. A–C, BMDMs were left untreated (CTRL), infected with WT Salmonella (MOI 10), or with the type III secretion system-deficient (SipB−) or flagellin-deficient (fljB−/fliC−) mutants for 1 h. Culture supernatants were collected and analyzed for secreted HMGB1 by Western blotting (A) and ELISA (B) and for secreted TNF-α by Luminex assay (C). D and E, BMDMs from WT and Ipaf−/− mice were left untreated (CTRL) or infected with WT Salmonella (MOI 10) for 1 h. Culture supernatants were collected and analyzed for secreted HMGB1 by Western blotting (D) and ELISA (E). F and G, BMDMs from WT and caspase 1−/− mice were left untreated (CTRL) or infected with WT Salmonella (MOI 10) for 1 h. Culture supernatants were analyzed for secreted HMGB1 by Western blotting (F) and ELISA (G). Black arrowhead on Western blots marks HMGB1 (29 kDa). ELISA and Luminex data represent the mean ± SD of triplicate samples from a single experiment, and all results are representative of three independent experiments.