Fig. 3.

Neutrophils regulate PCa growth in vitro. a LNCaP or C42B cell numbers were quantified by Trypan Blue Exclusion assay 24 h after direct co-culture with mouse bone marrow-derived neutrophils plated at a 10:1 ratio of neutrophil to cancer (n = 3). Asterisks denote statistical significance (*p < 0.05, **p < 0.01). b Caspase activity quantification during Incucyte S3 Live-Cell Analysis System imaging. Incucyte S3 Live-Cell Analysis System caspase-3/7 green reagent was added to neutrophil–PCa co-cultures when neutrophils were added to cancer cells. Quantification of cells was calculated based on presence of the green Caspase dye. Solid lines are PCa in culture; dashed lines are PCa cells cultured with neutrophils. Asterisks denote statistical significance (****p < 0.0001) (n = 3) for each condition. c Snapshot image of mouse neutrophils with cultured C42B cells imaged using NanoLive 3D Explorer for 1 h. White arrows denote neutrophils surrounding a C42B cell with membrane blebbing. Asterisks denote statistical significance (*p < 0.05, ***p < 0.001) d Trypan Blue quantitation of LNCaP and C42B 24 h after culture with human bone marrow neutrophils