Extended Data Fig. 6: CASP8(C362S) activates the ASC inflammasome in IECs.
From: Caspase-8 is the molecular switch for apoptosis, necroptosis and pyroptosis
a, Immunofluorescence confocal images of CASP8−/− HCT-116 clone 2 overexpressing either human wild-type caspase-8 or CASP8(C360S) together with DsRed–ASC untreated or treated with IDN-6556 (20 µM) and stained for caspase-8 after 24 h. Scale bar, 20 µm. Results are representative of two individual experiments. b, Immunoprecipitation of CASP8−/− HEK293T clone 1 lysates overexpressing either human wild-type caspase-8, CASP8(C360S) or empty vector (−) together with DsRed–ASC untreated or treated with IDN-6556 (20 µM) as indicated. Results are representative of two individual experiments. c, Immunofluorescence confocal images of BMDMs derived from Casp8fl/fl or Casp8C362S/fl mice treated with HTNCre for 24 h and stained with an anti-ASC antibody (top) or measurement of IL-1β levels in the supernatant of BMDMs (bottom; n = 3 biologically independent replicates). Scale bar, 20 µm. Dots and circles represent individual biological replicates. Data are mean ± s.e.m. One-way ANOVA followed by Sidak’s post-analysis compared to the corresponding untreated value. Results are representative of two individual experiments. d, ASC-speck-positive BMDMs (top; n = 100 of one representative experiment), measurements of IL-1β levels (middle; n = 3 biologically independent replicates) and LDH release (bottom; n = 3 biologically independent replicates) in the supernatants of BMDMs after treatment with LPS (200 ng ml−1), IDN-6556 (20 µM) or both (LPS and IDN-6556) for 24 h. Dots and circles represent individual biological replicates. Data are mean ± s.e.m. One-way ANOVA followed by Sidak’s post-analysis compared to the corresponding untreated value and shown for P > 0.1. Results are representative of two individual experiments.
