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. 2020 Sep 18;88(10):e00372-20. doi: 10.1128/IAI.00372-20

FIG 2.

FIG 2

LukAB contributes to bacterial survival when biofilms are in contact with neutrophils. (A and B) Wild-type USA300 biofilms were incubated with Cell Tracker Blue-labeled primary human neutrophils for 1 (A) and 2 (B) h, stained with Syto-9 (viable bacteria), and imaged using confocal laser scanning microscopy. Representative images of sections taken close to the base of biofilms. Insets show a ×1.2 optical zoom of the area outlined in white. (C and D) Experiments similar to those described for panels A and B, performed with an isogenic ΔlukAB USA300 strain. (E) Biofilm biomass measured at 1 h after incubation of wild-type (WT) and ΔlukAB USA300 biofilms with neutrophils for 1 h. (F) Total CFU per milliliter of biofilms from indicated strains, incubated with (+PMN) and without (−PMN) neutrophils for 1 h. Biofilms were grown in μ-slides and captured at a ×600 total magnification. Results are averages of six independent experiments performed in triplicate, with standard errors of the means (SEM). Images and measurements of volume were taken using Imaris software version X 6.4. Enumerations of CFU per milliliter were done independently, using biofilms grown in silicone tubing. Student t tests were performed for pairwise comparisons.