Fig 2. Screening of pneumococcal surface factors interacting with mucus.

Adherence of WT Spn and isogenic mutants to pooled mNL or hNF was assessed in a solid phase assay. Bacteria (2x 10⁴ CFU/100μl DMEM) were incubated with 100μL of undiluted, immobilized, pooled mNL in presence of 0.1% BSA for 2hr at 30°C. After 19 washes, adherent bacteria were determined by resuspending with 0.001% Triton X-100 following plating on TS agar plates supplemented with 200 μg/ml streptomycin. (A) Adherence of Type 4 (TIGR4), Δcps and Δcps::cps to mNF. (B) Adherence of Type 4 (TIGR4) and ΔrlrA to mNF. (C) Adherence of Type 23F and Type 23F::pilus-1 to hNF. Experiments were performed in duplicates and mean values of three independent experiments are shown with error bars corresponding to S.D. ****,p<0.0001 by one-way ANOVA followed by Dunnett’s multiple comparison test for multiple comparison (A) or Mann-Whitney test (B,C).