Fig. 5: Card9 inhibits NLRP3 inflammasome activation in a Ripk2-dependent manner.
From: Card9 protects sepsis by regulating Ripk2-mediated activation of NLRP3 inflammasome in macrophages
A Western blot analysis of interaction of Card9 and Ripk2 in BMDMs treated with LPS using co-immunoprecipitation (Co-IP). B Western blot of reciprocal Co-IP from HEK293T cells transfecting with Flag-tagged Card9 and HA-tagged Ripk2. C, D BMDMs were obtained from WT mice and Card9−/− mice and transfected with Ripk2 over-expression plasmid, cells were unstimulated (medium) or pretreatment with LPS (100 ng/ml) for 3 h, and then treated with ATP (5 mM, 1 h). C ELISA analysis of IL-1β and IL-18 secretion in the culture supernatant. D Western blot analysis of cell lysates. E, F BMDMs were transfected with Card9 over-expression plasmid, cells untreated or treated with Ripk2 inhibitor GSK2983559 (10 μM) for 1 h, followed by unstimulated (medium) or LPS + ATP treatment. E ELISA analysis of IL-1β and IL-18 secretion in the culture supernatant. F Western blot analysis of cell lysates. G Immunoblot analysis of Co-IP from BMDMs untreated or treated with Ripk2 inhibitor GSK2983559 (10 μM) for 1 h, followed by LPS treatment. Data are shown as mean ± SEM. Each group was representative of at least three biological replicates. *p < 0.05, **p < 0.01, ***p < 0.001.
