Activation of Nlrp3 Inflammasomes Enhances Macrophage Lipid-Deposition and Migration: Implication of a Novel Role of Inflammasome in Atherogenesis
Figure 7
ATP-induced lipid trafficking in lysosomes was blocked in Asc−/− BMMs.
Asc+/+ and Asc−/− BMMs were primed with low dose of 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). (A) BMMs were incubated with BSA-conjugated BODIPY FL-C5-lactosylceramide (LacCer) and lipid trafficking was tested by following LacCer trafficking. Representative confocal fluorescent images show the co-localization between LacCer (red color) and Lysotracker (green color). An increase in the yellow color in overlay images indicates increased LacCer trafficking to the lysosomes. (B) Quantified and summarized data showing the percent of lipid in lysosomes. (C) The ganglioside GM1 levels in isolated lysosomes from BMMs were determined by dot blot analysis. Represent dot blot image shows the ganglioside GM1 level in isolate lysosome homogenates as detected by cholera toxin-conjugated HRP. (D) Summarized analysis of ganglioside GM1 in lysosomes. * P<0.05 vs. untreated Asc+/+ control group; # P<0.05 vs. Asc+/+ ATP group (n = 6).