TRIM20 degrades inflammasome components through autophagy. (A) Levels of NLRP3 were determine in lysates from THP-1 cells subjected to TRIM20 or scrambled (Scr) siRNA were activated with 1,000 U/ml IFN-γ for 3 h, and 2.5 µg/ml LPS for 2 h (for optimal TRIM20 expression; Fig. S3 B). RI, relative intensity. (B) Levels of NLRP3 were determined in THP-1 subjected to TRIM20 or control knockdowns and treated or not with bafilomycin A1 (Baf A1). (C) The abundance of NLRP3 protein was determined in THP-1 cells subjected to TRIM20 or control knockdowns and exposed to Escherichia coli strain LF82 and IFN-γ in the presence or absence of bafilomycin A1. (D) The abundance of NLRP3 protein was determined in primary human MDMs subjected to TRIM20 or control knockdowns and exposed to LPS and IFN-γ in the presence or absence of bafilomycin A1. (E) Levels of NLRP3 were determined in THP-1 cells subjected to ULK1, Beclin 1, or scrambled (Scr) siRNA and treated with IFN-γ and LPS. (F–H) Levels of NLRP3 (F), NLRP1 (G), or pro–caspase 1 (H) were determined in cells expressing GFP or GFP-TRIM20 after autophagy induction (Earle's balanced salt solution , 3 h) in the presence or absence of bafilomycin A1. Data, means ± SE; n ≥ 3. *, P < 0.05; †, P ≥ 0.05 (ANOVA).
