Figure 3. Inhibition of DPP8/DPP9 elicits the inflammasome partially relying on ASC.
(A, B) ASC specks were imaged from B6Nlrp1b+ BMDMs treated with 10 μM VBP or 1G244 for 4 h or 8 h. (A) Representative confocal micrographs from three different experiments depicting DAPI (blue) and ASC (red). Arrows indicate ASC specks. (B) Quantification of the number of cells containing an ASC speck after 8 h of treatment with inhibitors are plotted as the mean ± SD obtained in each mosaic of three different experiments. (C, D) B6Nlrp1b+ or B6Nlrp1b+ASC−/− BMDMs were mock-treated or received 10 μM VBP (C) or 1G244 (D) in media containing SG and imaged on an IncuCyte platform. The number of positive cells was quantified relative to a Triton X100–treated well considered 100%. Values represent mean ± SD of technical duplicates of a representative experiment from three biological repeats. (F, G) B6Nlrp1b+ or B6Nlrp1b+ASC−/−BMDMs were treated with 10 μM of VBP (E, G) or 1G244 (F, G) for 8 h. Combined cell lysate and supernatant samples were immunoblotted for caspase-1 (E, F) and supernatants were probed for IL-1β (G) levels. Cytokine values represent mean ± SD of technical duplicates from three biological repeats. All data are representative from three biological repeats. Scale bars, 10 μm. ***P < 0.001; ****P < 0.0001 tested by one-way Anova with Tukey’s multiple comparisons.