Extended Data Figure 3: TcdB-induced caspase 1 inflammasome activation in Pyrin-complemented DC2.4 cells and human monocyte-derived macrophages.
From: Innate immune sensing of bacterial modifications of Rho GTPases by the Pyrin inflammasome
a–c, DC2.4 cells harbouring a vector (DC2.4+Vec), mouse Pyrin isoform 1 (DC2.4+mPyrin-iso1), mouse Pyrin isoform 2 (DC2.4+mPyrin-iso2), human Pyrin isoform 1 (DC2.4+hPyrin-iso1) or NLRP3 were stimulated with TcdB or as indicated. d–f, Phorbol-12-myristate-13-acetate (PMA)-differentiated THP-1 or U937 cells (harbouring a vector or human Pyrin isoform 1) were stimulated with TcdB or MxiH. g, qRT–PCR measurements of relative Pyrin expression level (normalized to that of GAPDH) (n = 3; mean ± s.d.). h, DC2.4 cells stably expressing eGFP–Pyrin were stimulated with TcdB or poly(dA:dT), and then subjected to anti-ASC immunofluorescence staining. DAPI stains the nuclei. The merged images show the co-aggregation of eGFP–Pyrin with endogenous ASC. Representative caspase 1 immunoblots from at least three repetitions are shown in c and d. ELISA assay of IL-1β release in a and percentages of cell death measured by LDH release in b, e and f are mean ± s.d. (n = 3).
