Fig 2. Human neutrophils do not require transcription or translation to release NETs in response to C.albicans.

(A, B) Inhibitors were used at the following concentrations: Actinomycin D (1 μg/ ml), flavopiridol (0.05 μM), CAS 577784-91-9 (10 μM), CHX (1 μg/ ml). Cells were infected with opsonized C.albicans at MOI 5 and subsequently fixed for immunofluorescence staining. (A) NET formation was quantified at 2h after infection by immunofluorescence staining of chromatin and Hoechst. Graphs show mean ± SEM from independent experiments with 3 different donors. (B) Representative images of an early time point (2h) of human neutrophils infected with C.albicans and treated with transcription or translation inhibitors. Cells were stained with PL2-3 (chromatin, red), NE (Neutrophil elastase, green) and Hoechst (blue). (C) Representative images of a late time point (4h) of C.albicans-infected neutrophils showing spread NETs that are unaffected by inhibitors of gene expression. (A) Statistical analysis revealed no significant changes after inhibitor treatment.