FIG 4.

Metabolic inhibition activates the NLRP1B inflammasome in murine macrophage cell line RAW264.7. (A) WT RAW264.7 cells were treated for 4 h with either medium alone, 10 μg/ml LPS, 250 mM 2DG, or a combination of LPS and 2DG. The cell lysates were probed for pro-caspase-1, caspase-1 p10, and β-actin by immunoblotting. (B) WT RAW264.7 cells and three NLRP1B KO clones from independent CRISPR constructs were treated with 10 μg/ml LPS and the indicated concentrations of 2DG for 4 h. The cell lysates were assayed for ATP (top) and were probed for pro-caspase-1, caspase-1 p10, and β-actin by immunoblotting (bottom). Blots are representative of three independent experiments. Graphed data represent means ± standard deviations from three independent experiments.