Extended Data Fig. 4. Additional data for RcsA and BaeR regulon overexpression, and effect of Spy overexpression on permeability.

a. Disk diffusion induction of plasmid-borne TAT-tagged TseH in bacterial lawns of E. coli BW25113 containing RcsA or vector only plasmids. IPTG was added throughout the lawn to induce RcsA. Disks contain arabinose to induce TAT-TseH expression. Representative of three independent replicates. b. Relative survival (killing by TseH^WT / TseH^H64A) of E. coli ΔdegP mutant or A. dhakensis with vector (Vec) or overexpressing genes from the BaeR regulon. One-way ANOVA with Sidak’s multiple comparisons test comparing samples to their respective empty vector controls; *, p < 0.05; **, p < 0.01; ns, not significant. Vector controls are shown in Figure 2 and are shown again here for comparison. Mean and standard deviations are shown. Dots show individual replicates. pSpy^ΔSec, plasmid expressing Spy with its Sec secretion tag deleted. c. Images of ΔbaeR E. coli overexpressing plasmid-borne TAT-TseH and either Spy or vector-only controls. The phase-contrast and propidium iodide(PI) channels are merged. Scale bar shows 5 μm. Representative of three independent replicates. d. Quantification showing the percent of total cells showing PI+ fluorescence. N number at top indicates total cells counted. Dots show mean results from three independent experiments (used to calculate statistics) and error bars show one standard deviation. Unpaired 2-tailed t-test, p = 0.0513.