Adherence of Type 23F::pilus-1 and isogenic mutants to pooled mNL and hNF was assessed in a solid phase assay. Bacteria (2 x 104 CFU/ 100 μl DMEM) were incubated with 100μL of undiluted, pooled mNL or 10 μg hNF 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 23F::pilus-1; nanB::janus and nanB::nanB to mNF (B) Adherence of Type 23F::pilus-1;
nanB::janus and ΔnanA,nanB::janus to hNF (C) Adherence of Type 23F::pilus-1; nanB::janus and NanBD270A to hNF (D) Adherence of Type 23F::pilus-1; ΔnanA and ΔnanA::nanA to mNF (E) Adherence of Type 23F::pilus-1; ΔnanA and ΔnanA::nanA to hNF. (A) A representative data-set is shown with error bars corresponding to S.D. ****,p<0.0001 by one-way ANOVA followed by Dunnett’s multiple comparison test. (B-E) Experiments were performed in duplicates and mean values of three independent experiments are shown with error bars corresponding to S.D. *,p<0.05; **,p<0.01; ***,p<0.001; ****,p<0.0001 by one-way ANOVA followed by Dunnett’s multiple comparison test for multiple comparisons.