“…PAM cells were seeded into 12-well culture plates (8×10 5 cells/well) at 0 h, stimulated by B.suis LPS at the concentrations of 0.1, 1, 10, 100, 1000 μg/mL after 24 h, harvested at 48 h. And then, the concentration of 100 ng/mL B. suis LPS was used for stimulation of the time points of 3, 6, 12, 24, 48 and 96 h. siRNAs transfection: PAM cells were seeded into 12-well culture plates (8×10 5 cells/well) at 0 h, to optimize siRNA transfection efficiency, we used X-tremeGENE siRNA Transfection Reagent (Roche, Basel, Switzerland) to transfect with the FITC-siRNA (Santa Cruz Biotechnology, CA, USA) at 12 h post-seeding, optimizing the ratio of X-tremeGENE siRNA Transfection Reagent and FITC-siRNA obtained a more than 90% transfection efficiency measured by flow cytometry. Caspase-11 siRNA targeting caspase-11 (Santa Cruz Biotechnology) siRNAs or Control siRNA-A (scramble siRNA, Santa Cruz Biotechnology) were introduced into PAM cells as described previously (Jiao et al, 2016), and according to the instructions of X-tremeGENE siRNA Transfection Reagent (Roche, Basel, Switzerland) .…”