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. 2015 Dec 11;5:17935. doi: 10.1038/srep17935

Figure 1. Genipin inhibits NLRP3 and NLRC4 inflammasome-mediated IL-1β secretion and caspase-1 activation in mouse macrophages.

Figure 1

LPS-primed BMDMs were incubated with genipin (200 μM unless otherwise indicated) or DMSO for 1 h, followed by treatment with various NLRP3 or NLRC4 inflammasome agonists. Culture supernatants were analyzed for IL-1β and TNF-α by ELISA. Precipitated cell supernatants (Sup) or cell extracts (Lysate) were immunoblotted using various antibodies. (A) IL-1β secretion in BMDMs stimulated with the indicated doses of genipin and ATP (5 mM, 1 h). (B) IL-1β secretion in BMDMs stimulated with genipin plus ATP, nigericin (Nige, 20 μM, 1 h), MSU (100 μg/ml, 4 h) or Listeria (MOI = 20, 4 h). (C) IL-1β secretion in BMDMs stimulated with the indicated doses of genipin and Salmonella (MOI = 20, 4 h). (D) IL-1β secretion in BMDMs transfected with 20 μg/ml flagellin or BSA for 8 h using Lipofectamine 2000. (E) TNF-α secretion in BMDMs stimulated with genipin plus ATP, nigericin, MSU, Listeria, Salmonella or flagellin. (F) Cell supernatants and cell extracts immunoblotted for caspase-1, IL-1β and ASC. GAPDH and β-tubulin served as loading controls. The data are representative of three independent experiments. **P < 0.01.