Cells were lysed with cell lysis buffer (Cell Signaling, 9803S) according to the manufacturer’s instructions. Rapid Gold BCA Protein Assay kit was used to determine protein concentrations in the cell lysate, and equal amounts of protein were separated by SDS–PAGE and transferred to a nitrocellulose membrane. Membranes were incubated overnight with primary antibodies against caspase-1 (AdipoGen, AG-20B-0042-C100, 1:5,000) and β-actin (MA5-15739, Invitrogen, 1:10,000). HRP-linked secondary antibody (Cell Signaling, 7076S, 1:3,000) and ECL reagent (Amersham, RPN2209) were used to visualize protein bands.
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