FIG 3.

HahK and HnoX are not required for pABA/calcium induced biofilms. (A) Colony biofilm formation was evaluated following growth on TPC of the following strains: WT (ES114), ΔhahK (KV7964), ΔhnoX (KV8025), and ΔhahK ΔhnoX (KV8484) strains. (B) Colony biofilm formation was evaluated following growth on TPC of the following strains: ΔsypF+sypF-hpt (KV7226) (same image as Fig. 2A), ΔhahK ΔsypF+sypF-hpt (KV9968), and ΔhnoX ΔsypF+sypF-hpt (KV10226). The data in this panel were obtained at the same time as those in Fig. 2A, and the ΔsypF+sypF-hpt is repeated here to facilitate comparisons across figures. Pictures were taken using the Zeiss Stemi 2000-c microscope (×6.5 magnification) at 72 h before and after disruption using a toothpick. Pictures are representative of three separate experiments. Arrows indicate where “pulling,” indicating cohesion, was observed. Dotted arrows represent where “pulling” was observed but to a lesser extent, with portions of the colony breaking off during disruption. (C) Biofilm cohesion was quantified from images as described in Materials and Methods with scores of 1 assigned to a null phenotype and 4 to the most/strongest. Statistics for panel C were performed via a one-way ANOVA using Tukey’s multiple-comparison test, where biofilm strength was the dependent variable. No significance was reached.