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. Author manuscript; available in PMC: 2017 Aug 25.
Published in final edited form as: Cell. 2016 Aug 25;166(5):1198–1214.e24. doi: 10.1016/j.cell.2016.07.027

Figure 1. ARIH1 Broadly Interacts with Components of Assembled, Neddylated CRLs.

Figure 1

(A) Left to right: immunoblot of low-level induced expression of FLAG-HA-ARIH1 or catalytic C375S mutant in 293 FLP-IN cells; high-confidence interacting proteins (HCIPs) for WT ARIH1 and ARIH1C357S, ± neddylation inhibitor MLN4924, identified by CompPASS (Sowa et al., 2009). ARIH1 predominantly interacts with CRL subunits colored by CRL1, blue; CRL2, yellow; CRL3, purple.

(B) TMT method to identify MLN4924-sensitive/neddylation-dependent ARIH1 HCIPs.

(C) ARIH1C357S HCIPs decreased ≥2-fold by neddylation inhibitor MLN4924 colored by CRL: CRL1, blue; CRL2, yellow; CRL3, violet. CRL3KLHL12 and substrate SEC13-SEC31A are highlighted. Error, ±SEM.

(D) Log2 fold change of ARIH1C357S interactors from the schematic in (C). Error bars, ±SE.