|
Fig. S9
The Gbp4-mediated resistance to S. Typhimurium is independent of IL-1β processing and pyroptotic cell death.
(A-E) Zebrafish one-cell embryos were injected with standard control (Std), Asc (A), Gbp4 (C), or Il1b (D, E) MOs in combination with antisense (As) or Gbp4 mRNAs (B) and zfiGluc mRNA (A-C), treated by immersion with vehicle alone (DMSO), 100 μM of a general inhibitor of caspases (PINH) or 100 μM of a specific inhibitor of caspase-1 (C1INH) (A), and infected at 2 dpi with ST (MOI of 10). The luciferase activity determined at 24 hpi as described in the Method section (A-C), while survival (D) and caspase-1 activity (E) were determined as described in Figures 2A and 2B, respectively. The Gluc was used as a positive control (C+). (F) Zebrafish lys:dsRED one-cell embryos were injected with standard control (Std) or Gbp4 MOs and infected at 2 dpf in the otic vesicle with ST at a MOI of 100. YO-PRO compound was injected at 3 hpi in the otic vesicle and pictures of each larva were taken at 4.5 hpi at the fluorescent microscope to visualize dead cells. Representative pictures are shown for each treatment. As a positive control, a group of cells dying during larvae development in the front part of the head are shown. The sample size for each treatment is 25 in A-C, 300 in D, 30 in E, 40 in F. S.I., ST infection. ns, not significant. **p<0.01; ***p<0.001 according to log rank test (D) or ANOVA followed by Tukey multiple range test (A, B, C and E).