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. 2016 Feb 12;23(7):1219–1231. doi: 10.1038/cdd.2015.176

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Copyright © 2016 Macmillan Publishers Limited

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Punicalagin retain mature IL-1β in response to different NLRP3 activators. (a) IL-1β ELISA in cell lysates (orange bars) or supernatants (black bars) from BMDMs primed with LPS (1 μg/ml, 4 h) followed by no stimulation (−) or stimulation with ATP (5 mM, 30 min), nigericin (10 μm, 30 min), hypotonic solution (90 mOsm, 1 h), monosodium urate crystal (MSU; 200 μg/ml, 3 h), melittin (5 μM, 30 min), or double-stranded DNA (dsDNA; 2 μg/ml, 30 min) in the absence or presence of punicalagin (PUN; 25 μM). (b) Extracellular LDH from macrophages treated as in (a). (c) IL-1β ELISA in supernatants (white bars) or extracellular LDH (black bars) from mouse bone marrow-isolated neutrophils primed with LPS (100 ng/ml, 4 h) followed by no stimulation (−) or stimulation with nigericin (10 μm, 30 min) in the absence or presence of punicalagin (PUN; 25 μM). (d) IL-1α ELISA in supernatants (left panel) or extracellular LDH (right panel) from BMDMs primed with LPS (1 μg/ml, 4 h) followed by no stimulation (−) or stimulation (+) with zVAD (100 μM, 20 h) in the absence or presence of punicalagin (PUN; 25 μM). (e) TNF-α and IL-6 ELISA in supernatants from BMDMs primed with LPS (1 μg/ml, 4 h), washed, and followed by incubation for 30 min in the absence or presence of punicalagin (PUN; 25 μM). *P<0.05; ***P<0.001 (Student's t-test)