Figure 4: ASC is critical for LeTx-induced speck formation. | Nature Communications

Figure 4: ASC is critical for LeTx-induced speck formation.

From: Activation of the NLRP1b inflammasome independently of ASC-mediated caspase-1 autoproteolysis and speck formation

Figure 4

(a) B6, B6Nlrp1b+ and B6Nlrp1b+ASC−/− BMDMs were grown on coverslips and treated with LeTx (10 μg PA and LF, respectively) for 60 min after which cells were fixed in 4% paraformaldehyde and immunostained for ASC (red) and caspase-1 (green). DAPI (blue) was included to localize nuclei. Arrows indicate specks. Confocal micrographs were acquired at × 60 magnification. (b) Quantification of specks per field. (c) B6, B6Nlrp1b+ and B6Nlrp1b+ASC−/− BMDMs that were stimulated with LeTx (high, 10 μg PA+10 μg LF; low, 500 ng PA+250 ng LF) for 30 min, and then incubated with the active caspase-1 Fam-YVAD-fmk FLICA probe for 45 min before fluorescent micrographs were acquired at × 20 magnification. * indicates magnified area. (d) Quantification of FLICA-positive cells/field and mean grey value. Data are representative of results from two independent experiments and quantification data are presented as mean±s.d. from a single representative experiment.

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