Figure 7.

Endocytosis was necessary in the process of DRibbles-inflammasome response. (a) CypHer5E-labeled DRibbles and lysotraker Green-labeled lysosome were collocated in THP-1 cells after 6 h incubation, and the nucleus were labeled with DAPI. (b) The endocytosis frequency of DRibbles by THP-1 cells was measured by FACS. DCs were pre-treated with ExoA (1 μM), Cytochalasin-D (Cyto-D, 1 μg/ml), and NH₄Cl (20 μM) for 30 min, then incubated with CypHer5E stained DRibbles (100 μg/ml) overnight. (c) Primed THP-1 cells were pre-treated with cytochalasin-D, ExoA, and NH₄Cl for 1 h, then stimulated with ATP/LPS (5 mM)/(10 ng/ml) and DRibbles (100 μg/ml) overnight. The supernatant IL-1β was detected by ELISA. (d) The frequency of IFN-γ⁺CD8⁺, IFN-γ⁺CD4⁺ T cells induced by pp65 DRibbles at indicated doses were measured by ICS, and (e) pp65 protein (10 μg/ml) and pp65 DRibbles (10 μg/ml). Results are shown as the means+S.D. and representative of two or more independent experiments. PBMC is from ⩾3 donors