Extended Data Fig. 7: ADAR1 deletion or Zα domain mutation induces ZBP1-mediated cell death of mouse fibroblasts.
From: ADAR1 prevents autoinflammation by suppressing spontaneous ZBP1 activation
a-j, Primary mouse lung fibroblasts isolated from mice of the indicated genotypes were stimulated with 200 U/mL IFN-α (a-c, e-j) or 200 U/mL IFN-γ (d) for 16 h or left untreated (ctrl). Next, cells were treated with 5 µg/mL CHX and 1 µg/mL anti-mouse TNF (a), 50 µM zVAD-fmk and 1 µg/mL anti-mouse TNF (b,c,g,h), 30 ng/mL mouse TNF and 10 µg/mL CHX or 50 µM zVAD-fmk (e,f,i,j) or infected with HSV1 ICP6mutRHIM at a multiplicity of infection of 5 (d). Cell death in (a,b,d,e,g,i) was quantified by measuring relative (rel.) SYTOX Green uptake every 2 h. c, Immunoblot related to (b). Samples were harvested 4 h post treatment. f,j Immunoblot related to (e,i). Samples were harvested 7 (TNF + CHX) and 3 (TNF + zVAD) hours, respectively. h, Immunoblot related to (g). Samples were harvested 8 h post treatment. Data points in (a,b,d,e,g,i) show the mean of 2-6 technical replicates (see Source Data) + SD and are representative of 3 independent experiments. Fitted lines represent a logistic growth fit. For gel source data, see Supplementary Figure 1
