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. 2018 Feb 1;8:16. doi: 10.3389/fcimb.2018.00016

Figure 5.

Figure 5

UPEC UTI89 colonization on fixed HTB9 uroepthelial cell layers under flow. (A) Graph showing the progression of bacterial growth on the surface by UTI89wt pre-cultured in LB (red line), UTI89wt pre-cultured in AU (green line) and UTI89ΔfimH pre-cultured in LB (blue line). The data points represent mean values of the bacterial coverage in percent at 20 min intervals from three separate flow chamber runs. Error bars represent ±1 SD between individual flow chamber runs (shown, for clarity, at 1-h intervals). Values at each 20 min time point obtained in each flow chamber run are mean values from recordings at 11 positions each representing 1,100 × 850 μm on the cell layer surface (in the graph, data is compiled from a total of 8,640 single scans). (B–D) Example of image data recorded at one representative 380 × 250 μm cropped position. Phase contrast (Bi–Biv,Ei–Eiv,Hi–Hiv), fluorescence microscopy (Ci–Civ,Fi–Fiv,Ii–Iiv), and software-detected coverage (Di–Div,Gi–Giv,Ji–Jiv) are shown at four specific time points. These are marked with broken lines in (A) representing different phases of surface colonization. Coverage by UTI89wt precultured in LB and AU differ significantly, at 5 and 9 h post seeding (p < 0.001, no overlap of the 95% CI). Coverage by UTI89wt and UTI89ΔfimH differ significantly at 5, 9, and 17 h post seeding (p < 0.001, no overlap of the 95% CI). GFP, green fluorescent protein; h, hours; PH, phase contrast microscopy; ROI, region of interest.