Figure 2. Lysosome rupture precedes cell death mediated by alum and LLOMe, but not by pyroptosis inducers. (A) Confocal microscopy of lysosome rupture in macrophages exposed to 250 ng/ml LPS in the presence or absence of 100 μM CA-074-Me for 2 h. Cells were then treated with 150 μg/ml alum, 2.5 mM LLOMe or 15 μM nigericin, and stained with Hoechst (blue), 20 MW FITC-Dextran (green), CellMask Orange (red), TO-PRO-3 (white). 2D-fluorescent intensity plots of FITC-dextran-labeled representative cells following alum, LLOMe and nigericin exposure. (B) Flow cytometry analysis of LLOMe and LT-treated cells. Murine macrophages were exposed to 2.5 mM LLOMe or 500 ng/ml LT, and lysosome and membrane integrity were measured at different time points using flow cytometry using LysoTracker and PI, respectively.