Fig. 5.

Constitutive activity of disease-associated mutant cryopyrin requires intact ATP-binding activity. (A and B) THP1 cells were transduced with recombinant retrovirus expressing the following proteins: empty vector (EV), no cryopyrin; WT cryopyrin; Walker A mutant (WA) cryopyrin; FCAS mutant cryopyrin (A439V); and FCAS mutant cryopyrin with Walker A mutation (A439V/WA). (A) IL-1β secretion was measured 12 h after transduction by ELISA. (B) IL-1β measured in A was corrected for the number of GFP⁺ cells in the culture. Error bars represent standard deviation between transductions performed in triplicate. (C) THP1 cells were transfected with plasmids expressing GFP and the following proteins: empty vector (EV), no additional protein; WT cryopyrin; FCAS/Muckle–Wells syndrome mutant cryopyrin (R260W); and R260W with Walker A mutation (R260W/WA). Secreted IL-1β was assayed by ELISA 24 h after transfection. Error bars represent standard error from duplicate measurements.