Figure 4. Physiological Role of NLRP6 Phase Separation in Anti-RNA Virus Defense in Hepatocytes.

WT, Nlrp6^−/− or Nlrp6^K350−354A knock-in mice were infected with MHV (i.p.) and analyzed as indicated (C–N).

(A) Immunoblot of NLRP6 in primary hepatocytes or intestinal epithelial cells (IECs) from Nlrp6-Flag knock-in mice.

(B) qPCR assays for Nlrp6 mRNA levels in different cells. N.D.: not detectable.

(C) Fluorescence immunohistochemistry staining of dsRNA (J2) and NLRP6 in liver sections from uninfected or MHV-infected WT and Nlrp6^−/− mice. Nuclei were stained with DAPI.

(D, E) Quantification of NLRP6-dsRNA (D) and NLRP6-ASC (E) puncta in liver sections. Data are mean ± SEM from 6 randomly sampled regions.

(F-H) Viral replication in the liver 3 days after MHV infection in Nlrp6^−/− (F), Asc^−/− (G) and Gsdmd^−/− mice (H) in comparison to WT mice.

(I, J) Serum (I) and liver-explant supernatant (J) IL-18 levels determined by ELISA from WT, Nlrp6^−/− and Asc^−/− mice 3 days post-infection.

(K) Fluorescence immunohistochemistry staining of dsRNA (J2) and NLRP6 in liver sections from uninfected or MHV-infected WT and Nlrp6^K350−354A mice. Nuclei were stained with DAPI.

(L, M) Serum (L) and liver-explant supernatant (M) IL-18 levels in WT and Nlrp6^K350−354A mice 3 days post-infection.

(N) TUNEL staining of MHV-infected WT and Nlrp6^K350−354A mice 3 days after infection. Scale bar: 20 μm.

See also Figure S5 and S6