Fig. 4: Piezo1 regulates neutrophil NET formation though SIRT2-HIF1α signalling during virus infection.

A C57BL/6 mice were infected with the PR8 virus for 48 h, and the lungs were collected. RNA was analysed via RNA sequencing to compare the expression profiles of control and virus-infected cells from the lung with those of certain genes involved in the ROS, inflammatory cytokine and chemokine signalling pathways. B–D Wild-type (WT) and Piezo1^−/− mice were infected with the PR8 virus for 48 h and treated with or without Yoda1 (2.6 mg/kg), as indicated. B mRNA expression of the indicated genes in neutrophils isolated from BALF. C Western blot of Piezo1 and SIRT2 in sorted neutrophils from BALF. D Intracellular staining of HIF1α in neutrophils isolated from BALF. E–G WT and Sirt2^−/− mice were infected with the PR8 virus for 48 h and treated with or without Yoda1 (2.6 mg/kg). E Expression of ROS in neutrophils isolated from BALF. Dot plots present representative data from flow cytometry analysis (left) and summarized data (right). F NET formation in neutrophils sorted from BALF by confocal fluorescence microscopy. Typical NET images are displayed (left), and the percentage of NETs was quantified (right). Scale bars, 50 µm; original magnification, 200X. G Western blot of PAD4 in sorted neutrophils from BALF. H, I WT and Hif1α^−/− mice were infected with the PR8 virus for 48 h and treated with or without Yoda1 (2.6 mg/kg). H Expression of ROS in neutrophils isolated from BALF by flow cytometry, and dot plots present representative data from flow cytometry analysis. I NET formation in neutrophils sorted from BALF by confocal fluorescence microscopy. Typical NET images are displayed (left), and the percentage of NETs was quantified (right). Scale bars, 50 µm; original magnification, 200X. The graph shows data from three independent experiments with four mice per group. **P < 0.01 and ***P < 0.001, compared with the indicated groups.