Fig. 3: Constitutive expression of the T3SS enabled vacuolar escape in 1 hour invasion assays.
From: Cross-membrane cooperation among bacteria can facilitate intracellular pathogenesis
A Mean GFP intensity of P. aeruginosa PAO1 with a GFP reporter plasmid for T3SS activity (pJNE05) when infecting human corneal epithelial cells (hTCEpi), comparing 1 and 3 h post-infection. B Representative widefield microscopy images of human corneal epithelial cells (hTCEpi) infected with P. aeruginosa wild type (WT) with a GFP reporter plasmid for T3SS activity using MOI 1. Images were taken at 1 and 3 hours postinfection at Ă—40 magnification. P. aeruginosa (green), Hoechst (gray). Scale bar equals 50 µm. C–E Image analysis of timelapse images measuring vacuole numbers (C), percent infected cells, represented as Area Under the Curve (AUC) bar plots (D), and cell death rates (E). F Representative widefield microscopy images of human corneal epithelial cells (hTCEpi) infected with P. aeruginosa wild type (WT), ΔexsA (T3SSOFF) or ΔexsE (T3SSON) strains at MOI 50 using 1 h invasion time. Images were taken at 10 hours postinfection at Ă—40 magnification. P. aeruginosa (green), Hoechst (gray). Scale bar equals 50 µm. Data of biological replicates represented as mean ± SD (A, D) or SEM (C, E), N = 3 (A), N = 8 (C, E), N = 6 (D). For statistical analysis, a two-sided Student’s t-test (A), a Two-way ANOVA with multiple comparisons (C, E), or a One-way ANOVA with multiple comparisons (D) was performed. Exact P values—(A) = 0.0216, (B): WT vs. ∆exsE = >0.0001, ∆exsA vs. ∆exsE = 0.0005. P ≤ 0.05 = *, P ≤ 0.001 = ***, P ≤ 0.0001 = ****. Source data are provided as a Source Data file.
