Fig. 4: Card9 over-expression decreases LPS-induced NLRP3 inflammasome activation in vitro.
From: Card9 protects sepsis by regulating Ripk2-mediated activation of NLRP3 inflammasome in macrophages
A, B BMDMs were transfected with Card9 over-expression plasmid. Cells were unstimulated (medium) or pretreatment with LPS (100 ng/ml) for 3 h, and then respectively treated with ATP (5 mM, 1 h), MSU (500 µg/ml, 2 h) or Nigericin (10 µM, 2 h). A ELISA analysis of IL-1β and IL-18 secretion in the culture supernatant. B Western blot analysis of BMDMs lysates. C–E BMDMs were transfected with Card9 over-expression plasmid, cells were unstimulated (medium) or pretreatment with LPS (100 ng/ml) for 3 h, and then stimulated with ATP (5 mM, 1 h). C Lysates of BMDMs immunoprecipitated with anti-ASC and then conducted immunoblot analysis. D, E Representative immunofluorescence images of ASC speck formation in BMDMs and quantification. Data are shown as mean ± SEM. Scale bars, 0.2μm. Each group was representative of at least three biological replicates. **p < 0.01, ***p < 0.001.
