Figure 2

E-protein suppresses NLRP3 inflammasome activation in bronchoalveolar lavage fluid (BALF). (A) Schematic representation of poly(I:C). WT and Nlrp3^−/− mice were injected intranasally with control and E-protein lentiviruses for 10 days. The mice were challenged intranasally with poly(I:C) for 24 h and bronchoalveolar fluid (BALF) and lungs were harvested to assess inflammasome activation. (B) Total white blood cells (WBCs) in BALF. WBCs were analyzed with Hematology Analyzer. Data are presented as mean ± SD, which were analyzed by one-way ANOVA coupled with Tukey’s test for multiple comparisons. (C) Quantification of LDH release into BALF. LDH release was assessed via LDH kit. Data are presented as mean ± SD, which were analyzed by one-way ANOVA coupled with Tukey’s test for multiple comparisons. (D) Quantification of IL-1β secretion into BALF. IL-1β secretion was assessed via IL-1β ELISA. Data are presented as mean ± SD, which were analyzed by one-way ANOVA coupled with Tukey’s test for multiple comparisons. ****P ≤ 0.0001, ***P ≤ 0.001, **P ≤ 0.01, *P ≤ 0.05. (E) Quantification of IL-18 secretion into BALF. IL-18 secretion was assessed via IL-18 ELISA. Data are presented as mean ± SD, which were analyzed by one-way ANOVA coupled with Tukey’s test for multiple comparisons. ****P ≤ 0.0001, ***P ≤ 0.001, **P ≤ 0.01, *P ≤ 0.05. (F) Western blot analysis of pro-IL-1β and NLRP3 in cellular lysates of WBCs in BALF. Data are presented as mean ± SD, which were analyzed by one-way ANOVA coupled with Tukey’s test for multiple comparisons. Blots were cut to probe with NLRP3 (from ~ 80 kDa to top) or pro-IL-1β and Actin (from ~ 80 kDa to bottom).