Fig. 4.

Mucin inhibition of biofilm formation by PAO1 is concentration dependent. PAO1/pMRP9-1 was grown aerobically on plastic coverslips in a 24-well microtiter plate in either LB or LBM broth containing variable concentrations of mucin for 24 h at 37 °C under aerobic conditions. CLSM micrographs and 3D image analysis of the PAO1 SABF grown (A) without mucin, and with concentrations of mucin indicated: (B) 10 mg/mL, (C) 5 mg/mL, (D) 2.5 mg mucin/mL, and (E) 0.625 mg/mL; magnification, 10×; bars, 200.00 nm. Images shown are representative of three independent experiments. (F) Twitching motility of PAO1 in different concentrations of mucin. PAO1 was stab-inoculated onto either LB or LBM (5 mg mucin/mL) twitching plates (1 % agar) containing variable concentrations of mucin. The plates were incubated at 37 °C for 48 h then at 23°C for an additional 48 h. The agar was removed and the twitching zones revealed by staining with crystal violet. Values represent the means of 3 independent experiments ± SEM. Twitching motility in decreasing concentrations of mucin was compared to that in 5 mg/ml mucin by one-way ANOVA; ***P<0.001.