Table 1.
Bacterial and eukaryotic cells and plasmids.
Fig 1.
Relative adhesion of FITC-labeled E. cloacae at different bacteria cell ratios (BCR) on cell lines from different tissues (T24 bladder cells, A-498 kidney cells, Caco-2 colon cells, A-431 epidermal cells and A-549 lung cells) as determined by flow cytometry.
A: BCR 100:1; B: BCR 10:1; C: 500:1. Relative data indicate the fluorescence related to uninfected cells. Values represent the mean ± SD from three independent assays and X-bar begins at Y = 1. *: p < 0.05, related to the relative adhesion of bacteria to T-24 cells.
Fig 2.
Confocal laser scanning microscopy of T24 bladder cells infected with E. cloacae at BCR 100:1.
Different z-stack sections reveal representative (A) top, (B) middle, and (C) below sections of T24 cells. The cross section in y axis view from the yellow line in A, B, and C images is represented in (D). E. cloacae cells are displayed in green after staining with FITC, cell nuclei of T24 cells (blue) are stained with DAPI, and wheat germ agglutinin stain host cell membranes with AlexaFluor™ 594 (red).
Fig 3.
Relative adhesion of FITC-labeled E. cloacae at BCR 500:1 to Caco-2, HT29 and HT29-MTX at different days of post-seeding (2, 10, and 25).
Relative data indicate the fluorescence related to uninfected cells and has been calculated from the median of the infected cells compared to that of the control without bacteria. Values represent the mean ± SD from three independent assays and X-bar begins at Y = 1.
Fig 4.
E. cloacae load isolated from the different cell lines after invasion assay at BCR 100:1.
Data indicate the CFU/mL isolated from T24, Caco-2, A431, A498, or A-549 cells. Values represent the mean ± SD from three independent assays. One-way ANOVA and Dunnett’s multiple comparison test (comparing all columns with T24) were used for statistical analysis. P < 0.05 was determined as statistically significant (*) and p < 0.01 as highly significant (**) compared to the bacterial invasion determined for T24 cells.
Fig 5.
Confocal laser scanning microscopy of T24 bladder cells infected with E. cloacae at BCR 100:1 for monitoring bacterial invasion into host cells.
Different z-stack sections reveal representative (A) top, (B) middle, and (C) below sections of T24 cells. The cross section in y axis view from the yellow line in A, B, and C images is represented in (D). E. cloacae cells are displayed in green after staining with FITC, cell nuclei of T24 cells (blue) are stained with DAPI, and wheat germ agglutinin stains host cell membranes with AlexaFluor™ 594 (red).
Fig 6.
Relative adhesion of E. cloacae to T-24 bladder cells (BCR 100:1, 60 min) observed in absence of fimA, fimH, or csgA genes, and complemented derivatives.
Data indicate the percentage of adhered bacteria in relation to the total bacterial count added to each sample (CFU/mL bacteria isolated after adhesion assay × 100 / CFU/mL bacteria added to each sample). NC: negative control (non-infected T-24 cells). Values represent the mean ± SD of three independent assays. **: p < 0.01, related to wild type bacteria.
Fig 7.
Relative adhesion and invasion results [%] of E. cloacae ATCC 13047 at BCR 100:1 against different cell lines, characterizing different tissue types.
Relative data are normalized against the interaction with T-24 bladder cells (= 100%). The degree of adhesion and invasion is colored by the heat map as displayed by the color bar legend. Identities closer to 0% are shown in red, and those higher than 100% in green.