Figure 6. NLRP3 is required for inflammasome activation downstream of TRIM21.

• A–C
  Primed HMDM were treated with MCC950 (1 μM) for 30 min before stimulation overnight with AdV (50,000 pp/cell) and h9C12 (20 μg/ml) or IVIg (20 mg/ml) or 10 μM Nigericin. IL‐1β (A) or TNF (B) or LDH release (C) was measured in cell supernatants. Data show average ± s.d. of two independent donors and are representative of three independent experiments.
• D
  WT, NLRP3, ASC or caspase‐1‐deficient THP‐1s were stimulated as in (A) and IL‐1β in the supernatant measured by ELISA. Data are representative of two independent experiments.
• E
  HMDM were primed with 10 μg/ml pI:C for 2 h, treated with 100 μM VX‐765 for 30 min before stimulation for 16 h as in (A). IL‐1β or LDH release was measured in cell supernatants. Data show average ± s.d. of two independent donors.
• F
  Primed HMDM were treated with KCl as indicated for 1 h, before being stimulated for a further 6 h as in (A). IL‐1β and TNF were measured in cell supernatants by ELISA. Data are representative of three independent donors.
• G
  Primed HMDM were stimulated with AdV and 20 μg/ml h9C12 or 10 μM Nigericin for 3 h and then immunostained for ASC or intracellular antibody. Co‐localisation was assessed by confocal microscopy. Data are representative of two independent donors.