MLN4924 inhibits NAE in human xenograft models of DLBCL. CB.17 SCID mice bearing OCI-Ly10 (A-D) and OCI-Ly19-luc (E-H) xenografts were administered a single subcutaneous dose of vehicle or MLN4924 at 10, 30, or 60 mg/kg. Tumors were excised at the indicated times and NEDD8-cullin conjugate levels (A,E), relative Nrf-2 protein levels (B,F), and relative pIκBα levels (C,G) were measured in the tumor lysates (20 μg protein per lane) by quantitative immunoblot analysis. Units are arbitrary. In panel F, the assay has a linear standard curve up to 750 units. Western blotting was also performed to analyze levels of cleaved caspase 3 after a single subcutaneous dose of 60 mg/kg (D,H). Vertical lines have been inserted to indicate repositioned gel lanes.