FIG 5.

The regulation of c-di-GMP synthesis and H1-T6SS activity by FleR is mediated by AmrZ. (A) Relative expression of amrZ measured by RT-qPCR in PAO1, ΔfleR, and ΔfleR(fleR) strains. (B) Relative expression of tse1 measured by RT-qPCR in the strains of PAO1 and ΔfleR with ectopic expression of amrZ. (C) Western blot analysis of Tse1-His in the cell-associated protein fractions from the strains of PAO1 and ΔfleR with ectopic expression of amrZ. (D) Bacterial killing assay between the indicated PAO1 strains and the E. coli prey. (E) Relative intracellular c-di-GMP levels in the indicated PAO1 strains were determined by measuring the relative expression of cdrA. Data are represented as the mean ± SD (n = 3). **, P < 0.01; ***, P < 0.001 versus the indicated group based on paired Student's t test. n.s., not significant; EV, empty vector for the control.