Protocol for building an in vivo model for observation and evaluation of neutrophil extracellular trapping nets in influenza virus-infected mice.

Neutrophils sense microbes and selectively release neutrophil extracellular traps (NETs) in response to pathogens. Here, we present a protocol for qualitative and quantitative analysis of NETs in PR8-infected mouse models. We describe steps for obtaining single-cell suspensions of infiltrated neutrophils from mouse bronchoalveolar lavage fluid (BALF). We then detail procedures to determine NET marker proteins using flow cytometry, confocal microscopes, and western blotting. This protocol may be applied to qualitative and quantitative analysis of NETs in influenza virus-infected mice. For complete details on the use and execution of this protocol, please refer to Zhang et al.1.