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. Author manuscript; available in PMC: 2013 Dec 1.
Published in final edited form as: Nat Immunol. 2013 May 5;14(6):543–553. doi: 10.1038/ni.2595

Figure 6. The NLRP3 inflammasome and caspase-1 regulate the pH of phagosomes containing Gram-positive but not Gram-negative bacteria.

Figure 6

a, Kinetics of caspase-1 activation in response to live S. aureus (MOI: 100) and E. coli (MOI: 100) was determined in WT and Ice−/− macrophage cell lines by immunoblot of Caspase-1 p10 in total cell lysates or cell supernatants. b, Kinetics of caspase-1 activation in response to S. aureus (red) and E. coli (grey) in BMDM was measured by flow cytometry (MOI: 50). c, YVAD-FLICA activity induced by uptake of live Gram-positive and Gram-negative bacteria (MOI: 50). Upper panel, Total YVAD-FLICA fluorescence; lower panel, flow cytometry histograms. FLICA was added at 15 min bacterial post-internalization for 1h. d-e Acidification of HI Gram-positive (d) and Gram-negative (e) bacteria-containing phagosomes in WT, Ice−/−, Nlrp3−/− and Asc−/− BM macrophage cell lines. NLRP3 inflammasome components (Nlrp3, Asc and caspase-1) are shown in green and blue. f, Total cellular ROS (H2DCFDA) induced by uptake of live Gram-positive or Gram-negative (MOI: 50) bacteria. ROS was measured at 15 minutes post-internalization. Data are representative of three independent experiments.