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. Author manuscript; available in PMC: 2018 Oct 25.
Published in final edited form as: Cell Rep. 2018 Aug 28;24(9):2356–2369.e5. doi: 10.1016/j.celrep.2018.07.098

Figure 6. Removal of Caspase-1 Inhibition by Negative Regulators Requires Calpain Activity.

Figure 6.

(A) ELISA analysis of IL-1β secretion in 293FT cells co-transfected with the indicated amount of plasmid (μg) for 24 hr.

(B) Immunoblotting analysis of Flightless-1 protein level change in BMDMs treated with the indicated stimuli (n = 5).

(C) Caspase-1-Flightless-1 interaction in BMDMs treated with the indicated stimuli was analyzed by immunoprecipitation with Caspase-1 antibody.

(D and E) Coomassie blue staining (D, upper panel) and silver staining (E) of proteins from BMDMs immunoprecipitated by Caspase-1 antibody. Two bands at around 230 and 45 kDa (arrows) were identified as myosin-9 and two species of actin by in-gel MS (D, lower panel).

(F) MS hit numbers of Caspase-1 interaction proteins in silica alone and silica + calpeptin treatment (1 hr) were analyzed by in-gel MS. Hits are grouped per functions. Flightless-1 is marked in red.

Data are representative of 2 or 3 independent experiments (A and C–E). Error bars indicate SEM. *p < 0.05, **p < 0.01, ***p < 0.001; ns, not significant (unpaired Student’s t test). See also Figures S5 and S6 and Table S1.