Figure 6.

GIT1 enhances affinity of NEMO for K63‐linked ubiquitin chains via CC2 of GIT1. A, HEK293T cells were transfected with Fag‐GIT1‐WT, HA‐NEMO‐WT, Myc‐ubiquitin‐WT, and Myc‐K6‐, K11‐, K27‐, K29‐, K33‐, K48‐ or K63‐linked‐Ub for 48 hours and subsequently treated with TNF‐α for 2 hours following MG132 (10 μM) treatment for 1 hour. HA‐immunoprecipitation was performed and analysed using anti‐Myc antibody by immunoblot analysis. B, HEK293T cells were transfected with HA‐NEMO‐WT, Myc‐ and K63‐linked‐Ub, and Flag‐GIT1, or Flag‐GIT1(ΔCC2) for 48 hours. Harvested protein was treated with TNF‐α for 2 hours following MG132 (10 μM) treatment for 1 hour, immunoprecipitated with anti‐HA and immunoblotted with anti‐Myc