Fig. 2.

Treatment of macrophages with VSF obtained from HNSCC cells reduces IL-1β and caspase-1 secretion. Peritoneal macrophages from wild-type C57BL/6 mice were initially treated overnight with VSF (1:5) (A) or VSF (1:5, 1:20, and 1:50) (B). Then the macrophages were primed with LPS (500 ng/mL) for 3h and stimulated with nigericin (10 μM) for 2h. Secretion of IL-1β was analyzed by ELISA. For the three analyzes, **p < 0.005 and ***p < 0.0005 when compared to control (EVs-free culture medium). Data are representative of four independent experiments with n = 3. (C) Protein expression of the active forms of IL-1β and caspase-1 in culture supernatant were evaluated by WB. β-actin was used as an endogenous control. Data are representative of three independent experiments with n = 3. Below each band, the numbers represent the respective densitometry quantification normalized in relation to β-actin expression. To determine the magnititude of reduction or enhancement of protein expression, positive controls were defined as 1.0 (100%) and were compared to VSF treatments.