Extended Data Fig. 6: Biofilm fitness quantification during mucosal colonization.

A. Fitness ratio based on the quantification of the total area measured from surface coverage by ∆bifA-mNeonGreen relative to WT-Scarlet during competition assays at the mucosal surface (HBE cultures). Each data point represents one imaged field of view (n = 15 for WT vs. WT; n = 12 for WT vs. ∆bifA) distributed within three biological replicates. The data for the WT-mNeonGreen vs. WT-mScarlet competition is the same from Fig. 3E but is shown here for comparison. Horizontal black lines mark the mean fitness ratio for each condition. B. Representative images of WT and ∆bifA competition assays in HBE cultures (maximum intensity projection; same field of view). For additional representative images, see Supplementary Fig. 1C. C. Cumulative distributions for cluster size formed by WT and ∆bifA. Distributions include three biological replicates. Vertical lines represent the median cluster-size for each strain. D. Percentage of small clusters (clusters smaller than 20 µm²) that comes from WT or ∆bifA populations. Each data point represents an independent biological replicate (n = 3); horizontal black lines mark their mean. Statistics: panel A and D, Welch unpaired t-test (two-sided) (*** p < 0.001, ** p < 0.01). All experiments were performed with CF HBE cells. Abbreviations: HBE cells, human bronchial epithelial cells; WT, wild-type. The data underlying this figure and information on statistical analyses can be found in the source data available with this manuscript (see Data Availability session).