Figure 6.

Baicalin suppressed nigericin-induced ASC speck formation and oligomerization. Bone marrow-derived macrophages were primed with LPS (500 ng/ml) for 4 h, pretreated with baicalin (0.6 mM) for 1 h, and triggered with nigericin (10 µM) for 1 h without LPS. (A) Representative immunofluorescence images showing ASC (red) and NOD-like receptor (NLR) family, pyrin containing domain 3 (NLRP3) (green) subcellular distribution. Nuclei (blue) were stained with Hoechst 33342. Merged images are also presented to show the co-localization of ASC specks and NLRP3 dots in the cytoplasm. Yellow arrows indicate ASC specks, and the enlarged cell showed with an ASC speck. Scale bars, 20 µm. (B) Percentages of cells containing ASC specks relative to the total number of cells in five randomly chosen fields with each containing ~50 cells. (C) Western blot analysis for ASC in Triton-X 100 insoluble pellets cross-linked with disuccinimidyl suberate. The experiments were performed three times independently, with one representative experiment shown. Data are shown as mean ± SD (n = 3). ***P < 0.001.