Figure 3.

Neutrophils migration characteristics in the presence of diverse pathogens

(A and B) The Fiji software plugin MTrackJ was used to track neutrophils. The orange box indicates the area imaged for all migration experiments and the blue box represents the area in which the neutrophils were tracked. Representative images with tracks are shown in B. Scale bar is 100 μm for A and B.

(C–F) Neutrophil migration parameters including speed (C), track length (D), total migration distance (E), and straightness (F) in response to P. aeruginosa, S. enterica, L. monocytogenes, or S. aureus from the cell tracks over 20-min intervals at 2-h time points. Data quantified from 14 lumens (P. aeruginosa), 12 lumens (S. enterica), 13 lumens (L. monocytogenes), or 14 lumens (S. aureus) across 5 independent experiments. Error bars represent the mean plus SEM. All bacteria were compared to each other at each time point and analyzed with ANOVA. For each condition, emmeans and SEM were calculated and pairwise comparisons were performed with Tukey’s adjustment. Asterisks represent significance of neutrophil migration for each bacterial species condition compared to L. monocytogenes condition at that time point. P values are labeled as ∗p < 0.05; ∗∗p <0 .001; #p < 0.0001. Individual data points are displayed with each gray scale color representing a different replicate.