Extended Data Fig. 5: AIM2 acts as an upstream regulator of RhoA modifications, and the ZBP1 Zα2 domain is required for cell death.
From: AIM2 forms a complex with pyrin and ZBP1 to drive PANoptosis and host defence
a, RhoA-GTP activity in wild type (WT), Aim2−/−, Mefv−/− or Zbp1−/− bone marrow-derived macrophages (BMDMs) infected with HSV1 or treated with TcdB for 12 h. Activity was normalized to total RhoA levels. Data are mean ± s.e.m. from three independent experiments. ns, not significant; ***P < 0.001; ****P < 0.0001 (one-way ANOVA with Dunnett’s multiple comparisons test; n = 3, 6, 7 or 9). b, c, Activated RhoA (RhoA-GTP) assessed using a pull-down assay with Rhotekin-RBD beads from WT, Aim2−/−, Asc−/− or Casp1−/− BMDMs infected with HSV1 (b) or F. novicida (c). Data are representative of at least three independent experiments. d, RhoA-GTP activity in WT BMDMs infected with HSV1 or transfected with poly(dA:dT) for 12 h. Data are mean ± s.e.m. from three independent experiments. ns, not significant; **P < 0.01 (one-way ANOVA with Dunnett’s multiple comparisons test; n = 3). e, Activated RhoA (RhoA-GTP) assessed using a pull-down assay with Rhotekin-RBD beads from WT, Aim2−/−, Asc−/− or Casp1−/− BMDMs transfected with poly(dA:dT). Data are representative of at least three independent experiments. f, g Cell death in WT, Zbp1−/−, Zbp1∆Za2/∆Za2 or Ripk3−/− BMDMs after HSV1 (f) or F. novicida (g) infections. Red indicates dead cells. Images are representative of at least three independent experiments. Scale bar, 50 μm. h, i, Quantification of the cell death from f (h) or g (i). Data are mean ± s.e.m. ****P < 0.0001 (one-way ANOVA with Dunnett’s multiple comparisons test; n = 9 from 3 biologically independent samples). Exact P values are presented in Supplementary Table 1. For gel source data, see Supplementary Figure 1.
