Fig. 5: RIPK3 cleavage modulates IL-1β production in vitro.

BMDMs were primed for 3 h with 25 ng ml⁻¹ LPS (a, c) or other TLR ligands (b) with the following concentrations: 5 μg ml⁻¹ poly(I:C); 2 μg ml⁻¹ P₃Cys; 2 μg ml⁻¹ CpG; 100 ng ml⁻¹ TNF. With the exception of time/dose-dependent compound A treatments in (a), BMDMs were treated for 5 h post-priming using 500 nM compound A (b) or 100 nM TAK1 inhibitor (c). Treatment using nigericin (NG; 5 μM) post-priming in (c) was 45 min. Cell supernatants were assayed for IL-1β by ELISA after treatment with indicated compounds. Data are represented as mean + SEM of N = 3 independent biological replicates per genotype. Graphs are representative of 3 independent experiments performed with 3 independent biological replicates each time. *P < 0.05, **P < 0.01, ***P < 0.005, *****P < 0.0005, ns not statistically significant. d Western blot of BMDMs upon 25 ng ml⁻¹ LPS time-course. e BMDMs were primed for 3 h either with 25 ng ml⁻¹ LPS or 5 μg ml⁻¹ poly(I:C), followed by 5 h of 500 nM compound A. Cell lysates were analysed by western blot. d, e Results are representative of 3 independent experiments performed with 3 independent biological replicates.