Extended Data Fig. 3. Additional E. coli mutants susceptible to T6SS-delivered TseH.

a. Disk diffusion induction of plasmid-borne TAT-tagged TseH expression in bacterial lawns of E. coli MG1655. Wild-type (top) or H64A mutant (bottom) TseH are compared. Right disks contain the inducer of expression, arabinose (Ara), and left disks contain the repressor, glucose (Glu). Representative of three independent replicates. b. As in a, comparing induction of plasmid-borne TAT-tagged TseH^WT expression in E. coli MG1655 or A. dhakensis (ΔT6SS). Similar data for TseH^WT in E. coli has been shown previously and is shown again here for comparison⁴. c. Relative survival (killing by TseH^WT / TseH^H64A) of E. coli wild-type or knockout mutants from indicated categories. One-way ANOVA with Dunnett’s multiple comparisons test comparing each sample to WT; **, p < 0.01; ***, p < 0.001; ns, not significant. Data for WT (E. coli) is shown in Figure 2 and is shown again here for comparison. Aux., auxotrophs. d. RT-qPCR comparing expression of genes in wild-type E. coli prey after T6SS competition assay with TseH^WT or TseH^H64A killer strains. Value with TseH^H64A killer was set to 1 to show fold induction in response to TseH activity upon delivery by the T6SS. One-way ANOVA (with samples shown in Figure 2d) with Sidak’s multiple comparisons test; ns, not significant. For both graphs, the mean and standard deviation are shown. Dots show individual replicates. TseH^WT, V. cholerae V52 with all anti-bacterial effectors inactivated except TseH; TseH^H64A, V52 with all anti-bacterial effectors inactivated including TseH.