Fig. 7: Bacterial cooperation occurred in vivo.
From: Cross-membrane cooperation among bacteria can facilitate intracellular pathogenesis
A Representative in vivo confocal microscopy images of murine corneas (ROSAmT/mG mice, tdTomato cell membranes, red) infected with different combinations of fluorescent (green) and non-fluorescent bacteria (ΔexsA-GFP + 10% ΔexsA, 90% ΔexsA-GFP + 10% ΔexsE, 10% ΔexsE-GFP). Images were taken 20 hours post-infection using the murine scratch-infection model, imaging infection foci within the corneal epithelium using a 60x water-immersion objective. Top panel shows en-face view of a single z-slice, middle panel shows en-face view of max intensity projections; bottom panel shows the corresponding lateral view as STD-projection. Scale bar equals 30 µm. B Histogram with LOWESS curve of bacterial GFP-signal distribution using different combinations of fluorescent and non-fluorescent bacteria (ΔexsA-GFP + 10% ΔexsA, 90% ΔexsA-GFP + 10% ΔexsE, 10% ΔexsE-GFP) normalized to max GFP signal for each image. C Bar plots quantifying percent cells infected with GFP-producing bacteria under the three different infection conditions (10% ΔexsE, 90% ΔexsA-GFP + 10% ΔexsE, ΔexsA-GFP + 10% ΔexsA). D Bar plot quantifying the average size of intracellular GFP-aggregates (particles over 5 μm3) in the three infection conditions (10% ΔexsE, 90% ΔexsA-GFP + 10% ΔexsE, ΔexsA-GFP + 10% ΔexsA). Data of biological replicates represented as mean ± SD, N = 3 (C, D). For statistical analysis, a one-way ANOVA with multiple comparisons (C, D) was performed. Exact P values—A: 10% ∆exsE GFP vs. 90% ∆exsA GFP ( + 10% ∆exsA) = 0.0092, 90% ∆exsA GFP ( + 10% ∆exsE) vs. 90% ∆exsAGFP (+10% ∆exsA) = 0.0131; B: 10% ∆exsE GFP vs. 90% ∆exsA GFP ( + 10% ∆exsE) = 0.0452, 10% ∆exsE GFP vs. 90% ∆exsA GFP ( + 10% ∆exsA) = 0.0014, 90% ∆exsA GFP ( + 10% ∆exsE) vs. 90% ∆exsA GFP ( + 10% ∆exsA) = 0.0305. P ≤ 0.05 = *, P ≤ 0.01 = **. Source data are provided as a Source Data file.
