Figure 3
Pasteurella multocida-induced pulmonary epithelial cell apoptosis contributes to the loss of pulmonary epithelial integrity. A Top ten RNA-seq enrichment pathways of cellular processes in murine lungs infected with 104 CFU of PmCQ2 at 40 hpi. B Western blot analysis of murine lungs infected with 104 CFU of PmCQ2 at different time points. Each lane represents one individual. C Representative images of confocal analysis of SPC staining and TUNEL staining in in situ murine lungs infected with 104 CFU of PmCQ2 or saline at 16 hpi. Scale bar = 20 μm. The red arrow indicates an apoptotic lung epithelial cell. D Representative images of TUNEL staining of murine lungs infected with 104 CFU of PmCQ2 at different time points. Scale bar = 200 μm. At the bottom are quantifications of TUNEL-positive cells in 3 random sections. The data are presented as the means ± SDs. E Representative images of TUNEL staining of murine lungs infected with 104 CFU of PmCQ2 or 107 CFU of PmCQ6 at 24 hpi. Scale bar = 50 μm. On the right, quantification of TUNEL-positive cells in 3 random sections is shown. The data are presented as the means ± SDs. F A schematic of the Transwell assay protocol. On the right, quantification of the number of translocated bacteria in TC-1 cells treated with 20 μM of the apoptosis inhibitor Ac-DEVD-CHO or saline at 8 hpi. Each group contained 3 replicates. The data are presented as the means ± SDs. * p < 0.05, *** p < 0.001.