Fig. S2

Sept15 and Sept7b morphants show increased susceptibility to S. flexneri infection.

A. Immunostaining of zebrafish larvae at 3 dpf with antibody against SEPT7 (red) in cells of the caudal fin epithelium (a), a neutrophil (mpx:GFP labeled) (b), and a macrophage (mpeg1:YFP labeled) (c). Scale bars, 10 μm. B. Survival curves of Ctrl or Sept15 morphants infected with T3SS- S. flexneri (ΔmxiD strain, low dose). Pooled data from 3 independent experiments per treatment using at least 15 larvae per experiment. Significance testing performed by Log Rank test. C. Enumeration of bacteria at 0, 24, or 48 hpi from Ctrl (open circles) or Sept15 (closed circles) morphants infected with T3SS- S. flexneri (ΔmxiD strain). Circles represent individual larvae, and only larvae having survived the infection (thus far) included here (i.e., dead larvae not homogenised for counts). Half-filled circles represent enumerations from larvae at time 0 and are representative of inoculums for both conditions. Pooled data from 3 independent experiments using up to 3 larvae per treatment. Mean ± SEM also shown (horizontal bars). Significance testing performed by Student’s t test. D. Representative western blot of extracts from larvae injected with Ctrl or Sept7b morpholino (Mo) using antibodies against GAPDH (as control) or SEPT7. E. Survival curves of Ctrl or Sept7b morphants infected in the HBV with S. flexneri M90T (low dose). Pooled data from 3 independent experiments per treatment using at least 15 larvae per treatment. Significance testing performed by Log Rank test. ***, P<0.001. F. Enumeration of bacteria at 0, 24, or 48 hpi from Ctrl (open circles) or Sept7b (closed circles) morphants infected with S. flexneri M90T (low dose). Half-filled circles represent enumerations from larvae at time 0 and are representative of inoculums for both conditions. Pooled data from 3 independent experiments using up to 3 larvae per treatment. Circles represent individual larvae, and only larvae having survived the infection (thus far) included here (i.e. dead larvae not homogenised for counts). Mean ± SEM also shown (horizontal bars). Significance testing performed by Student’s t test. *, P<0.05. G. Survival curves of Ctrl or Sept15 morphants infected in the caudal vein with S. flexneri M90T (low dose). Pooled data from 3 or more independent experiments per treatment using at least 15 larvae per experiment. Significance testing performed by Log Rank test. **, P<0.01. H. Enumeration of bacteria at 0, 24, or 48 hpi from Ctrl (open circles) or Sept15 (closed circles) morphants infected with S. flexneri M90T in the caudal vein. Circles represent individual larvae, and only larvae having survived the infection (thus far) included here (i.e., dead larvae not homogenised for counts). Half-filled circles represent enumerations from larvae at time 0 and are representative of inoculums for both conditions. Pooled data from 3 independent experiments using up to 3 larvae per treatment. Mean ± SEM also shown (horizontal bars). Significance testing performed by Student’s t test. **, P<0.01.