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. 2015 Feb 20;11(1):166–182. doi: 10.4161/15548627.2014.981915

Figure 3.

Figure 3.

Production of mitochondrial reactive oxygen following P. aeruginosa infection leads to inflammasome activation. (A) Flow cytometry of untreated BMDMs (basal) or infected as indicated and stained with MitoSOX Red. (B) Immunoblot as in Figure 1A of BMDMs pretreated for 1 h with Mito-TEMPO (250, or 500 μM) or NAC (10 or 25 mM) then infected with PA103ΔUΔT (MOI 25) for 4 h. The lower panels show ELISA of IL1B and TNF secretion as in Figure 1A. ** and *** indicate significant differences between the levels in the presence and absence of the Mito-TEMPO and NAC, P < 0.01 and < 0.001 respectively. (C) Flow cytometry of uninfected BMDM (basal) or pretreated for 1 h with Mito-TEMPO (500 μM) or NAC (25 mM) then infected with PA103ΔUΔT (MOI 25) for 4 h and stained with MitoSOX Red. (D) as in (C), but in cells infected with PAO1. (E) As (B) but in cells infected with PA01 as shown. (F) Flow cytometry of cells from Vav1-atg7−/− or Vav1-atg7+/+ mice left uninfected (basal) or infected with PA103ΔUΔT (MOI 25) for 4 h and stained with MitoSOX Red. (G) Immunoblot and ELISA as in (B) from Vav1-atg7+/+ and Vav1-atg7−/− BMDMs treated as shown. *** indicates significant differences from the levels produced from infected Vav1-atg7−/− cells, P < 0.001. n.s. not significant. All data are representative of 3 independent experiments.