Fig. 6. Baicalin significantly inhibits not only NLRP3 inflammasome activation but also the necrosis in macrophages treated with MSU crystals.

BMDMs from WT mice were primed with LPS (1 μg/mL) for 4 h, and then pre-treated without or with graded doses of baicalin for 1 h, followed by stimulation with MSU crystals (300 μg/mL) for 6 h in the presence or absence of baicalin. a The expression levels of indicated proteins in cell lysates and culture supernatants (Sup) were analyzed by Western blotting. β-Actin was adopted as a loading control for cell lysates. b The levels of secreted IL-1β in culture supernatants were determined by cytometric bead array (CBA). c Western blotting analysis of ASC oligomers in Triton X-100 insoluble pellets cross-linked with disuccinimidyl suberate. d Lytic cell death was measured by lactic acid dehydrogenase (LDH) release in culture supernatants (n = 5). e Representative images showing PI (red, staining dead cells) combined with Hoechst 33342 (blue, staining all cells) fluorescence. Scale bars, 20 μm. f Quantification of PI-positive cells in 5 randomly chosen fields and ratio of PI-positive over all cells (revealed by Hoechst 33342) is defined as the percentages of lytic cell death. Data are shown as mean± SD (n = 5). *P < 0.05; **P < 0.01; ***P < 0.001, NS not significant.