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. 2022 Mar 9;10(2):e02211-21. doi: 10.1128/spectrum.02211-21

FIG 3.

FIG 3

AmiL influenced pyocyanin and rhamnolipid production, elastase activity, and biofilm formation of P. aeruginosa. (A) PAO1 carrying the pROp200 empty vector (EV) or pROp200-amiL (AmiL+), and amiL-deleted PAO1 carrying pROp200 (△AmiL) were cultured in LB for 6 h, and AmiL expression was measured by qRT-PCR. (B) Growth curve analysis of the EV, △AmiL, and AmiL+ strains. (C) Pyocyanin in the supernatant of EV, △AmiL, and AmiL+ strains that cultured in LB for 24 h was detected. (D) The activity of elastase in the supernatant of EV, △AmiL, and AmiL+ strains cultured in LB for 8 h was measured. (E) Biofilm formation of the indicated strains that cultured in LB in the 96-well plate for 24 h was determined. (F) The indicated AmiL-constructed strains were cultured in M9 medium for 16 h, and rhamnolipid in the supernatant was measured. (G) The EV, △AmiL, and AmiL+ strains were cultured in LB for 24 h, and Staphylococcus aureus was used to perform the LasA staphylolytic assays. (H) Exotoxin A activity of the indicated strains cultured in Trypticase soy broth (TSB) with 1% glycerol for 24 h was measured. Data are shown as mean ± SEM of at least three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, non-significant.