Supplementary Figure 1: LUBAC mediates recruitment of the non-canonical kinases TBK1 and IKKε to and their activation in the TNFR1-SC.
From: TBK1 and IKKε prevent TNF-induced cell death by RIPK1 phosphorylation
(a) A549 HOIP Control or HOIP KO cells were stimulated with TNF (200 ng/mL) for the indicated times and subjected to lysis, followed by western blot analysis. (b) A549 HOIP Control or HOIP KO cells were stimulated with FLAG-TNF (500 ng/mL) for the indicated times. Lysates and TNFR1-SC were analysed by western blot. (c) HOIP-deficient HeLa cells reconstituted with empty vector, HOIPWT or enzymatically inactive HOIPC885S were stimulated with FLAG-TNF (1 µg/mL) for the indicated time. Lysates and TNFR1-SC were analysed by western blot. (d) HOIP-deficient A549 cells reconstituted with empty vector, HOIPWT or enzymatically inactive HOIPC885S were stimulated with FLAG-TNF (1 µg/mL) for the indicated time. Lysates and TNFR1-SC were analysed by western blot. Unprocessed original scans of blots are shown in Supplementary Figure 7. (a-d) One experiment representative of two independent experiments is shown.
