Skip to main content
. Author manuscript; available in PMC: 2014 Nov 15.
Published in final edited form as: J Immunol. 2013 Oct 2;191(10):10.4049/jimmunol.1301490. doi: 10.4049/jimmunol.1301490

Figure 6. Mitochondrial ROS is required for NLRP3 inflammasome dependent events.

Figure 6

(A) WT BMDM were LPS primed, treated with nigericin following YVAD exposure, and labeled with MitoTracker Red and LysoTracker Green. Cells imaged via live cell microscopy for 1 hr. Data presented as positive cells per field at 60 min. Error bars represent ± s.d. n=5. WT BMDM were LPS primed and treated with nigericin following 100 µM YVAD, or 100 µM or 500 µM MitoTEMPO (Mito T). Supernatants were analyzed for (B) IL-1β via ELISA or (C) LDH release as percentage of LDH release by Triton-X 100 treated control cells. Error bars represent ± s.d. n=3 (D) WT BMDM were LPS primed, nigericin and 500 µM MitoTEMPO treated. LysoTracker and MitoTracker (not shown) MFI recorded via live cell microscopy. The LysoTracker MFI was normalized to initial signal and background corrected. LysoTracker data are presented as one line/cell from five representative fields and split into two groups: cells that lost MitoTracker signal after 60 minutes of imaging (top panel) and cells that retained MitoTracker signal throughout the experiment (lower panel). (E) WT BMDM were LPS primed and nigericin or ATP treated with or without 500 µM MitoTEMPO. Cells were labeled with MitoTracker Red and imaged via live cell microscopy. Data are presented as percent of cells/field retaining MitoTracker signal. Error bars represent ±s.d. n≥ 10 fields. (**, P=0.0061; ***, P< 0.0001).