Figure 3.

The presence of NEMO or NEMO-A323P prevents RIPK3 from integrating FADD-containing complex II. (a) Endogenous complex I pull-down was performed by Flag-TNF immunoprecipitation in KO, NEMO and A323P-NEMO cells upon different periods of Flag-TNF stimulation (min). Immunoblot analysis for the indicated proteins is shown. (b and c) KO, NEMO and NEMO-A323P cells were stimulated with TNF (10 ng/ml) for the indicated periods (min). Complex II was isolated by an FADD immunoprecipitation. Immunoblot analysis was performed to reveal the indicated proteins. Single asterisk indicates modification of RIPK1 when caspases are active and double asterisks indicate cleaved form of Caspase-8. A representative immunoblot revealing the levels of the indicated proteins is shown. IP, immunoprecipitation; TCL, total cell lysate