Fig. 8

Caspa is required for the Caiap-mediated resistance to S. Typhimurium. (A,B) Zebrafish one-cell embryos were injected with standard control (Std) or Caiap MOs in combination with antisense (As) or Caspa mRNAs. At 2 dpf, embryos were infected with ST and survival (A) and caspase-1 activity (B) were determined as described in Figures 5A,B, respectively. S.I., ST infection. (C–F) HEK293T cells were transfected with zebrafish Caiap-mCherry or CaiapΔCARD-mCherry in the presence or absence of zebrafish wild-type FLAG-Caspa or catalytic inactive FLAG-Caspa (C230A), fixed at 48 h post-transfection and imaged using a laser confocal microscope (C,D), or lysed and immunoprecipitated with ANTI-FLAG M2 Affinity Gel and probed with antibodies to FLAG and mCherry (E) or lysed and resolved under non-reducing and reducing conditions (F). Representative views of maximum-intensity projection images of HEK293T cells are shown in (C), quantitation of the percentage of Caiap specks in relation to the total number of Caiap transfected cells is shown in (D) and representative blots are shown in (E,F). The sample size for each treatment is 300 for (A), 30 for (B), and 50 for (C). Scale bar, 5 µm; not significant; *p < 0.05; **p < 0.01; ***p < 0.001.