Fig. 3
Membrane lipid rafts mediated the binding, endocytosis, oligomerization and activity of βγ-CAT. a Frog peritoneal cells were treated with MβCD and then incubated with βγ-CAT. The detergent-resistant membrane fractions were extracted by density gradient centrifugation, and the distribution of βγ-CAT on membrane lipid rafts was determined by western blotting. Flotillin-1 is a marker of lipid rafts. The immunoblots are representative of three independent experiments, the original images of immunoblots are shown in Supplementary Figure 9. b Frog peritoneal cells were treated with MβCD and then incubated with FITC-labeled βγ-CAT. The binding of βγ-CAT to peritoneal cells was evaluated by flow cytometry. The untreated normal frog peritoneal cells are indicated as normal, and frog peritoneal cells treated with FITC-labeled βγ-CAT are indicated as control. c The colocalization of βγ-CAT and membrane lipid rafts was observed by confocal microscopy. Scale bar = 10 μm. d Frog peritoneal cells were treated with MβCD and then incubated with βγ-CAT. The oligomerization of βγ-CAT was detected by western blotting. The immunoblots are representative of three independent experiments, the original images of immunoblots are shown in Supplementary Figure 10. e The caspase-1 activation and IL-1β release induced by βγ-CAT was measured by western blotting. The immunoblots are representative of three independent experiments, the original images of immunoblots are shown in Supplementary Figure 11
