Fig. 2: The conjugation ability of IncX3 plasmid, and the effect of ΔvirBR on colonisation, adhesion and pili formation.

a The in-vitro conjugation frequency of 3R, 3R-ΔvirBR, 3R-ΔvirBR-pUC19-virBR, 3R-ΔvirBR-pUC19 strains. Data are means ± SEM; n = 3 biologically independent replicates obtained from three independent experiments. One-way ANOVA and Tukey’s multiple comparisons test were performed on values. b Abundance in chicken cecum of 3R (circles and black bars) and 3R-ΔvirBR group (squares and red bars) over time (days). n = 3 biologically independent replicates. c Abundance in mice faeces of 3R (circles and black bars) and 3R-ΔvirBR group (squares and red bars) over time (days). The detail experimental design can be found in Fig. S10. n ≥ 3 biologically independent replicates obtained from three independent experiments. Groups were compared using unpaired two-tailed t-test. d The adhesion ability to Caco-2 cells of 3R-ΔvirBR and 3R-ΔvirBR-pUC19-virBR (complemented with intact virBR) compared with 3R parent strain (given as 100%). Data are means ± SEM; n = 3 biologically independent replicates obtained from three independent experiments. One-way ANOVA and Dunnett’s multiple comparisons test were performed on values. e The adhesion ability to IEC-6 cells of 3R-ΔvirBR and 3R-ΔvirBR-pUC19-virBR (complemented with intact virBR) compared with 3R parent strain (given as 100%). Data are means ± SEM; n = 3 biologically independent replicates obtained from three independent experiments. One-way ANOVA and Dunnett’s multiple comparisons test were performed on values. f. Transmission electron micrographs illustrating pili formation of 3R, 3R-ΔvirBR, 3R-ΔvirBR-pUC19-virBR. Scale bars, from left to right - 2 μm, 500 nm and 200 nm. Image is representative of three independent experiments. Source data are provided as a Source Data file.