Figure 6
OmpAAb induces mitochondrial fragmentation in vivo. (A) Illustration of the mouse infection (created with BioRender.com). (B) Neutropenic BALB/c mice were infected intranasally with 107 colony forming units (CFUs) of the indicated bacteria. Six hours post-infection bronchoalveolar lavage (BAL) fluid was collected from the trachea of mice. Alveolar macrophages were isolated and immunofluorescence was performed using anti-TOM20 antibody to stain mitochondria (red) and DAPI to stain nuclei. Scale bar represents 10 µm. (C) Quantification of cells with fragmented mitochondria. The cells displaying a discontinuous segmented mitochondrial network were scored as fragmented. The quantification was performed in a blinded manner. Error bars represent standard deviation, n = 157–521 cells. One-way ANOVA with Tukey’s multiple comparisons test *p \(\le \) 0.05, **p \(\le \) 0.01. (D) Immunofluorescence was performed on the alveolar macrophages isolated from the BAL fluid from mice infected with ΔOmpA + OmpAAb::Flag complemented strain using anti-TOM20 antibody to stain mitochondria (red), anti-Flag antibody to stain OmpA::Flag (green) and DAPI to stain the nucleus (blue). Arrows indicate OmpAAb::Flag (green) colocalization with mitochondria (red). Scale bar represents 10 µm.
