Fig. 6

ROS-mediated cell death and S100A12 release depend on gasdermin D (GSDMD). Healthy control neutrophils (5 × 10⁶/ml) were stained with SytoxGreen and incubated for 4 h a without or b with PMA (100 nM) stimulation. Where indicated, inhibitors DPI (10 μM), C23 (30 μM), canakinumab (10 μg/ml), and anakinra (200 ng/ml) were used 1 h prior to culture. Cell death was quantified as amount of fluorescence at 523 nm. Supernatants were harvested and measured for S100A12 protein content by ELISA (c) and for LDH using the LDH assay kit according to the manufacturer’s protocol (d). Data are mean and SD of 3–8 independent individuals (b). Statistical significance was calculated Brown-Forsythe and Welch’s ANOVA test followed by Dunn’s multi-comparison test (* = p > 0.05, ** = 0.005, *** = p > 0.0005)