For exogenous gene expression in HEK293T and HeLa, cells were transfected with EZ-Trans cell Transfection Reagent (Life-iLab) according to the manufacturer’s instructions. For gene expression in macrophages, cells were transduced with recombinant lentivirus prepared by transiently transfecting lentiviral vectors (pCDH-puromycin, Δ8.91 and VSV-G) expressing WT or mutants of ASC into HEK293T cells for 48 h60. Transduced macrophages were selected with 5 μg/mL of puromycin for 7 days and protein expression was validated by immunoblotting.
