Figure 3.

Elevated expression of A20 enhanced ripoptosome formation but impaired TNF complex I formation. (A,B,D) HaCaT cells were transduced with A20 LV. A20 expression was induced by 4-HT, and the cells were stimulated as indicated. (A) Caspase-8 immunoprecipitation (IP) and (B) TNF complex I precipitation were performed as described in the Materials and Methods. Recruited proteins were analyzed by WB. (C) Two A20 CRISPR HaCaT clones were treated as indicated, and TNF complex I precipitation was performed as described in the Materials and Methods. (D) TNF complex I and caspase-8 complexes were sequentially precipitated from the same cellular lysates and analyzed by WB.