Extended Data Fig. 4 ∣. CQ31 selectively activates the CARD8 inflammasome.

(a) Viability of cell lines after treatment with CQ31 for 24 h as assessed by CTF. (b) Human resting T-cells were treated with CQ31 (10 μM) or VbP (10 μM) for 24 h before assessing cell viability by CTG and pyroptotic and apoptotic markers by immunoblotting. Data is representative of two independent experiments. (c) N/TERT-1 immortalized keratinocyte cells were treated with CQ31 (10 μM) or VbP (10 μM), incubated for 1 h, and stained with PI. PI uptake was assessed over 14 h. (d) HEKa immortalized keratinocyte cells were treated with CQ31 (10 μM) or VbP (10 μM), before assaying for LDH release. (e) HEK 293 T cells stably expressing CASP1 and GSDMD were transfected with plasmids expressing NLRP1 and ASC and treated with CQ31 (10 μM) or VbP (10 μM) for 24 h. GSDMD cleavage was assessed by immunoblotting. (f) RAW264.7 cells were treated with CQ31 (100 μM) or VbP (2 μM) for 24 h, before assaying for LDH release. (n = 3) (g) J774.1 cells were treated with various doses of CQ31 before cell viability was measured by CTG and CTF. Data in a (n = 4), b,d,f (n = 3), and c,g (n = 6) are means ± SEM of the indicated replicates. All data except where indicated, including immunoblots, are representative of three or more independent experiments.