Recombinant vectors encoding mouse Flightless-1 with a C-terminal myc tag and LRRFIP2 with a C-terminal HA tag were constructed by PCR-based amplification of complementary DNA from mouse BMDMs. Truncated mutants of BCAP were generated by PCR-based mutation and amplification of WT BCAP expression vector (3) using Q5 Site-Directed Mutagenesis Kit (E0554S, New England Biolabs). Expression plasmids encoding mouse ASC (pUNO1-mASC, InvivoGen), NLRP3 (puno1ha-mnalp3, InvivoGen), RFP–caspase-1 (C285A) (43), LRRFIP2, BCAP, and Flightless-1 were transfected into HEK293T cells with TransIT-LT1 transfection reagent (MIR 2300, Mirus Bio) according to the manufacturer’s instructions.

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