Figure 4.

Comparison of neutrophil responses to C. glabrata during biofilm and planktonic growth. (a) Planktonic and biofilm C. glabrata were co-cultured with human neutrophils at an effector:target ratio of 1:2 for 4 h and fungal inhibition was estimated by an XTT assay. Results were normalized to the no neutrophil controls, and data from 3 experiments performed in triplicate were combined. Statistical significance was determined using a two-tailed Student’s t-test assuming unequal variances, *P < 0.05, SEM shown. (b) Neutrophils were co-cultured with C. glabrata for 4 h and NET release was estimated by Sytox Green staining of free DNA at various time points. Results were normalized for the positive control, PMA, and data from 4 experiments performed in triplicate were combined. The statistical significance for NET release to biofilm and planktonic C. glabrata were analyzed for each time point using a two-tailed Student’s t-test assuming unequal variances, *P < 0.05, SEM shown. (c) NETs were induced by incubation with PMA for 1.5 h prior to addition to C. glabrata. After 10 min, fungal inhibition was estimated by an XTT assay. Results were normalized to the no neutrophil controls, and data from 4 experiments performed in triplicate were combined. Statistical significance was determined using a two-tailed Student’s t-test assuming unequal variances, *P < 0.05, SEM shown.