Figure 4
A. baumannii infection perturbs mitochondrial physiology in an OmpAAb dependent manner. (A) A549 cells were infected with the indicated bacteria for 6 h at MOI 50. The cells were treated with the CellROX reagent to examine ROS (green) and DAPI to label nuclei (blue). 30% (v/v) H2O2 treatment used at a final concentration of (1:1000) for 1 h at 37 °C served as positive control. Scale bar represents 20 µm. (B) The bar graph represents the quantification of ROS levels in A549 cells after the indicated infection/treatment. The experiment was done in triplicates, n = 90–103 cells. Error bars represent standard deviation. One-way ANOVA with Tukey’s multiple comparisons test ****p \(\le \) 0.0001. (C) A549 cells were infected with the indicated bacteria for 6 h at MOI 50. The cells were treated with the CellROX reagent to examine ROS (green) and DAPI to label nuclei (blue). Scale bar represents 20 µm. (D) The bar graph represents the quantification of ROS levels in A549 cells after the indicated infection. The experiment was done in triplicates, n = 72–85 cells. Error bars represent standard deviation. One-way ANOVA with Tukey’s multiple comparisons test ****p \(\le \) 0.0001. (E) TMRE assay to examine mitochondrial membrane potential in A549 cells after 24 h of infection with the indicated bacteria. The experiment was done in triplicates, Error bars represent standard deviation. One-way ANOVA with Tukey’s multiple comparisons test ****p \(\le \) 0.0001. (F) ATP levels in A549 cells after 24 h of infection with the indicated bacteria. The experiment was done in triplicates. Error bars represent standard deviation. One-way ANOVA with Tukey’s multiple comparisons test **p \(\le \) 0.01, ****p \(\le \) 0.0001. All the experiments shown here were performed three times independently.
