Encephalomyocarditis Virus Viroporin 2B Activates NLRP3 Inflammasome
Figure 5
Mitochondrial ROS- and cathepsin B-independent activation of NLRP3 inflammasome by EMCV.
(A) LPS-primed BMMs were infected with EMCV. Cell-free supernatants were collected at indicated time points and analyzed for IL-1β by ELISA (left y axis, filled bars). Cells were collected at indicated time points and stained with MitoSOX for 30 min and analyzed by flow cytometry. The proportions of ROS-producing mitochondria are shown (right y axis, open circles). (B–C) LPS-primed BMMs were treated with EMCV, ATP, MSU (B), or Alum (C) in the presence or absence of Mito-TEMPO (500 µM), a scavenger specific for mitochondrial ROS, or cathepsin B inhibitor CA-074 Me (10 µM) (C). Cell-free supernatants were collected at 24 h (EMCV, Alum) or 6 h (ATP) post infection or stimulation, and analyzed for IL-1β by ELISA. Data are representative of at least three independent experiments, and indicate the mean ± S.D. *, P<0.05.