Fig. 3.

Cdt-induces HMGB1 release from macrophages. Panel A: THP-1^WT-derived macrophages were incubated with Cdt containing CdtB^WT, CdtB^R117A or CdtB^A163R Following incubation for 24 hrs, supernatants were harvested and analyzed by Western blot for the presence of HMGB1. Results are plotted as HMGB1 levels (% of levels released by control cells; 0 Cdt) versus Cdt concentration and CdtB subunit type; results represent the mean±SEM of four experiments. A representative Western blot is shown for HGMB1. Panel B: Macrophages derived from THP-1^WT , THP-1^Casp1−, THP-1^Casp4− and THP-1^Syk− cells incubated with medium (solid bars) or 50 ng/ml Cdt (hatched bars); 24 hrs later, supernatants were harvested and analyzed for the presence of HMGB1. Results are plotted as HMGB1 release (% of release observed in THP-1^WT- derived macrophages incubated with Cdt) vs cell type. Results are the mean±SEM of three experiments. * indicates statistical significance (p<0.05) when compared to control cells incubated in medium alone (panel A) or to THP-1^WT- derived macrophages incubated with Cdt. A representative full blot is shown in Fig. 5S.