Supplementary Video S2 from Low-Dose Ionizing γ-Radiation Elicits the Extrusion of Neutrophil Extracellular Traps

Supplementary Video S2. Time-lapse confocal microscopy of co-cultures of tumor spheroids covered by radiation-induced NETs and cytotoxic lymphocytes. HT29 tumor cell-spheroids stained with CMRA (red) were cocultured or not with 2 Gy-irradiated neutrophils in the presence/absence of DNAse I to allow the presence of NETs (Sytox Green, green) around the spheroids. Cytotoxic immune cells (T or NK lymphocytes, blue) were pre-stained and cocultured with tumor spheroids for 4 hours before confocal microscopy acquisition. Time-lapse videos show a representative z-slice of the co-cultures over time (8 hours, 2,400× accelerated). As quantified in Figures 2 A, B and D, E, kinetically monitoring the tracks of the lymphocytes, data show evidence for the avoidance of the NETs in the migration paths.