Extended Data Fig. 9: Immunofluorescence imaging-based opsonization assays of A. baumannii A74.
From: Uncovering bacterial pseudaminylation with pan-specific antibody tools
(A) Density plots of biological replicates assessing the intracellular number of A. baumannii A74 in THP-1 macrophages treated in the absence of added antibody, or in the presence of Pse 1E8 or a mouse IgG1 isotype control with >200 cell assessed for each biological replicate. (B) Violin plots depicting the abundance of intracellular bacteria in THP-1 macrophages treated with A. baumannii A74 in the absence of added antibody, or in the presence of Pse 1E8 or a mouse IgG1 isotype control. Statistical analysis corresponding to pairwise two-sided student t-tests with p-values less then 0.001 denoted by *** and corresponding to p-values = 2.88×10−36 and 9.32×10−29. Statistical analysis was undertaken on the combined data from four biological replicates corresponding to n > 900 cells per group. The boxplot denotes the upper (Q3) and lower (Q1) quartiles of the observed data ranges, with the median denoted between these ranges. The whiskers denote 1.5 times the interquartile Range of Q1 and Q3, excluding outliers. (C) A representative immunofluorescence microscopy micrograph of THP-1 cells one hour after infection with A. baumannii A74. DNA has been stained with Hoechst 33342 (blue), actin with SiR-Actin (red), and Pse mAb 1E8 (green). Scale bar is 5 μm.
