Fig. 4.

NLRP12-mediated inhibition of NF-κB and ERK dampens cytokine production and clearance of Salmonella in macrophages. (A) BMDMs from WT and Nlrp12^−/− mice were treated with Salmonella LPS. Nlrp12^−/− BMDMs were also treated with either the ERK inhibitor U0126 or the IKK2 inhibitor SC-514 before and during LPS stimulation. Cell lysates were collected and analyzed for P-ERK, ERK, P-IκBα, and IκBα by Western blotting. (B–D) WT and Nlrp12^−/− BMDMs were treated with SC-514 during S. typhimurium infection. (B) Cell lysates collected after 2 and 24 h of infection were plated for bacterial colony-forming units (CFU). (C) Culture supernatant from BMDMs collected after 16 h of Salmonella infection was analyzed for IL-6 and KC by ELISA. (D) Nitrite in the culture supernatant collected after 24 h of infection was measured with Griess reagents. Data represent the mean ± SD of triplicate wells. Data are representative of at least three independent experiments. *P < 0.05.