Effect of CIAS1 mutation on the induction of cell death and ASC-dependent NF-κB activation. (A) 106 THP-1 cells were transfected with 0.5 μg of an expression vector for GFP-tagged CIAS1 wild-type (WT), CIAS1 SNP S196N, or one of the disease-associated mutants of CIAS1 (R260W, L264F, D303N, G307S, Y441H, N477K, E567K, Y570C, M662T, and G755R), and incubated with PMA (10 ng/mL) for 4 hours. The percentage of dead cells (7-AAD-positive) among the population of GFP-positive cells is shown. Data represent the means (± SD) of triplicate determinations, and are representative of 3 independent experiments. (B) HEK293FT cells were transfected with 16 ng of an expression vector for CIAS1, or one of its mutants, in the presence or absence of 16 ng of an expression vector for ASC. The induction of NF-κB is shown as fold-change compared with cells that were transfected with a control vector without ASC (set equal to one). Values are the means (± SD) of triplicate determinations, and data are representative of 2 independent experiments.
