Research Paper Volume 12, Issue 2 pp 1747—1759

K63 ubiquitin chains target NLRP3 inflammasome for autophagic degradation in ox-LDL-stimulated THP-1 macrophages

Figure 4. P62 mediates the autophagic regulation of NLRP3 inflammasomes. (A) The expression of p62 in the lysates of Mφ, which were left untransfected or transfected with vehicle, p62-siRNA, or control siRNA for 48 hours. (B) The immunoblot analysis of the lysates of Mφ, which were left untransfected or transfected with vehicle, p62-siRNA, or control siRNA for 24 hours, and subsequently treated with or without rapamycin, and stimulated with ox-LDLs (50 ug/ml) for another 24 hours. Rapamycin was administered to cells for one hour prior to ox-LDL stimulation. (B and C) The densitometric analysis of the NLRP3, ASC, (B) and p20 (C) signal, which were normalized to β-actin. (D) The ELISA of IL-1β in the supernatants obtained from (B). The data are presented as mean ± SD (n=3); * denotes the statistical significance by one-way ANOVA with post hoc Dunnett’s multiple comparisons test. *P<0.05, **P<0.01, ***P<0.001.