Extended Data Fig. 8: Binding on additional anti-Kp mAbs and ROI definition and spot detection for mAb binding characterization, related to Fig. 3.

(A) Scatter plot displaying the Mean Fluorescence Intensity (MFI) at the single bacterium level of the mAb binding against ST147_NDM-1 c.i. 1 (black lines show median with interquartile range). Median values ± SD are the following: 1419 ± 976 a.u., n = 28503 bacteria over 2 independent experiments for 08D18; 1246 ± 643 a.u., n = 29643 bacteria over 2 independent experiments for 08K19; 731 ± 450 a.u., n = 27873 bacteria over 2 independent experiments for 05M13; 1189 ± 561 a.u., n = 29066 over 2 independent experiments bacteria for 08F04; 1034 ± 524 a.u., n = 25337 bacteria over 2 independent experiments for 05C11; 118.2 ± 1.46 a.u., n = 29118 bacteria over 2 independent experiments for unrelated. One-way ANOVA with Tukey’s multiple comparisons with 95% C.I. to compare each anti-Kp mAb to the unrelated mAb resulted in the following P values, **** p < 0.0001. (B) Images show ST147_NDM-1 c.i. 1 stained with 08D18, 08K19, 05M13, 08F04, 05C11 mAbs labelled with anti-human A488 conjugated secondary antibody (green). Bacterial DNA is stained with DAPI (blue). Experiments were repeated two times obtaining similar results. Scale bar 2 µm. (C) Images on the left show the binding pattern of some A488-labelled anti-Kp mAbs on ST147_NDM-1 c.i. 1 bacteria. Images in the middle show the ROI where A488 signal was detected whereas on the right it is shown the morphology of A488 spot. Experiments were repeated four times obtaining similar results. Scale bar 2 µm.