Necroptosis blockage improves the disease outcome in KPn pneumonia.

(A). C57BL/6 mice infected intranasally with 3.0x104 CFUs of KPn received intraperitoneally 100μl of 100μM necrostatin-1s or vehicle (DMSO) 2hrs prior to infection and then every 4hrs for 12hrs post-infection. Mice were sacrificed at 3 days post infection, lungs were isolated and were processed for flow cytometry analysis of neutrophils by staining with anti-Ly6G-APC and anti-CD11b-Pacific Blue antibodies. Representative dot plots from one out of 3 independent experiments is shown. (A’) shows percent neutrophils (Ly6G+CD11b+) as mean ± SEM from three independent experiments with 3–5 mice per group in each experiment. Statistical analysis was done by ANOVA with Dunn’s post hoc analysis (**, p(B). In separate set of experiments performed similarly as in (A), lungs were homogenized aseptically and plated to enumerate bacterial burden. Each dot represents one mouse. n = 9 in each group in 3 independent experiments. (***, p<0.001).