Figure 4: WEHI-345 specifically blocks cytokine production on NOD stimulation.

(a,b) Wild-type BMDMs were treated with IFNγ+MDP (10 μg ml⁻¹), TNF (100 ng ml⁻¹), Smac-mimetic (compound A, 500 nM) or LPS (2.5 ng ml⁻¹). Cytokine levels of TNF (a, n=6–9) and IL-6 (b, n=3–6) were determined after 24 h by enzyme-linked immunosorbent assay (ELISA). Values were normalized to the average of control samples for better comparison. Absolute values of control samples for TNF (a) were: MDP: 142 pg ml⁻¹, TNF: 29 pg ml⁻¹, cA: 446 pg ml⁻¹, LPS: 3.9 ng ml⁻¹) and for IL-6 (b): MDP: 29 pg ml⁻¹, TNF: 61 pg ml⁻¹, cA: 188 pg ml⁻¹, LPS: 12.9 ng ml⁻¹). (c) Inflammasomes in LPS-primed wild-type BMDMs were activated using Alum (250 μg ml⁻¹), ATP (5 mM), Smac-mimetic (Compound A, 500 nM) or Staurosporine (1 μM), and levels of IL-1β in the supernatant was measured by ELISA. Values were normalized to the average of control samples (Alum: 508 pg ml⁻¹, ATP: 220 pg ml⁻¹, compound A: 230 pg ml⁻¹, Staurosporine: 124 pg ml⁻¹). n=3–4. In all panels, * indicate P values <0.05 and ** indicate P values <0.005 based on a non-paired Student’s t-test. Error bars are s.e.m.