Fig. 5.

HNSCC-derived VSF inhibits the induction of inflammasomes components. Peritoneal macrophages (5 × 10 (Yang et al., 2011) cells/well for ELISA and 1 × 10 (Zhang et al., 2015) for WB assays) from wild-type C57BL/6 mice were initially treated with the HNSCC-derived VSF (1:20) overnight. Then the macrophages were primed with 500 ng/mL LPS for 3h and stimulated with 10 μM nigericin for 2h. (A) The secretion of IL-6 was evaluated by ELISA. (B) Protein expression of the precursor forms of IL-1β and caspase-1 in cell lysates were analyzed by WB. β-actin was used as endogenous control. 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. (C) NLRP3 gene expression in VSF-treated macrophages in comparison to non-treated cells was determined by RT-PCR. All values were normalized using the β-actin as an endogenous control. Data are representative of independent experiments with n = 3. **p < 0.005 compared to the control.