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. 2014 Jan 27;9(1):e87552. doi: 10.1371/journal.pone.0087552

Figure 6. Asc gene deletion and caspase-1 inhibition blocked cholesterol deposition in lysosomes.

Figure 6

BMMs were primed with LPS (1 ng/ml) for 3 h and treated with ATP (2.5 mM, 16 h) in the absence or presence of caspase-1 inhibitor WEHD (0.15 µg/ml), or IL1β (0.5 ng/ml) alone. Then BMMs were loaded with oxLDL (10 µg/mL) for 16 hours. (A) Representative confocal fluorescent images showing the colocalization between filipin (cholesterol) and Lamp1 (lysosome marker). An increase in the purple color in overlay images indicates increased cholesterol trapping in lysosomes. (B) Quantified and summarized data showing co-localization co-efficiency between filipin and Lamp1. (C) Effect of Asc gene deletion on cholesterol level in isolated lysosomes from BMMs. Lysosomes were isolated from Asc+/+ and Asc−/− BMMs and cholesterol concentration in these isolated lysosomes were determined using a standard fluorescence assay kit. Some groups of cells were pretreated with ATP in the presence of IL1R antagonist (IL1Ra, 40 ng/ml) or IL18 (25 nM) alone. * P<0.05 vs. untreated Asc+/+ control group; # P<0.05 vs. Asc+/+ ATP group (n = 6).