*2.5. Induction of Neutrophil Extracellular Trap (NET) Formation with Pseudomonas aeruginosa*

Isolated neutrophils, 25,000/well, were incubated with *P. aeruginosa* MOI 10 in a glass-bottom 96-well plate (MatTek Corporation, Ashland, MA, USA) precoated with poly-D-lysine 1 mg/mL (Sigma-Aldrich/Merck, Darmstadt, Germany) for 1 or 4 h at 37 ◦C and 5% CO2, and sterile medium was used as a negative control. As we did not observe any significant difference in NET formation in control conditions (in the absence of *P. aeruginosa)* between 1 and 4 h (see Supplementary Figure S1B), the control values for 4 h were used for the further analysis.

To study the effect of G-CSF on the capacity of neutrophils to form NETs, neutrophils isolated from the blood of healthy volunteers (*n* = 5) were challenged with *P. aeruginosa* MOI 10 in the absence or presence of human G-CSF (Filgrastim HEXAL, Holzkirchen, Germany) at a concentration 10 ng/mL for 4 h.
