Figure 7.

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