Fig. 3.

HNSCC-VSF does not induce cell death. For the viability assays, peritoneal macrophages from wild-type C57BL/6 mice were treated overnight with VSF or EVs-free medium followed by priming with LPS (500 ng/mL) for 3h and stimulation with nigericin (10 μM) for 120 min (A and C) or 70 min (B). (A) Frequency of positive cells for ethidium bromide (Et/Br) was determined by fluorescence microscopy. (B) Incorporation of Sytox Green by permeable cells was determined in a Incucyte Zoom microscope. (C) The LDH release was measured using commercial kits, according to the manufacturer's instructions ***p < 0,0001 in comparison to the control. Data are representative of three independent experiments with n = 3. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)