FIGURE 5.

Transwell assay to assess the protective capacity of a M. catarrhalis catalase mutant. Dual species S. pneumoniae (SpnEF3030) and NTHi 86-028 NP (NTHi86028) biofilms were seeded in 24-well plates on a fixed H292 cell monolayer. To the transwell inserts, approximately 10⁵ CFU per mL M. catarrhalis 7169 (solid), Mcat7169 ΔkatASpec1 (vertical stripes) or CDM (open) was added. Protection was assessed at (A) 24 h and statistical analysis was performed using a one-way ANOVA and Dunnett’s multiple comparison test of log transformed CFU per mL (B) and 48 h and statistical analysis was performed on log transformed CFU per mL using the Kruskal–Wallis with Dunn’s multiple comparison test. Bars represent the mean and standard deviation of three independent assays with a minimum of two technical replicates. P-value of <0.01 denoted ^∗∗ and P-value of <0.001 denoted ^∗∗∗.