Figure 4
From: Disordered region of cereblon is required for efficient degradation by proteolysis-targeting chimera
CRBN degradation by VHL-CRBN heterodimerizing PROTACs recapitulates a CRBN deficiency. (A) Hep3B cells were glutamine-starved and treated with TD-158 (500 nM) for 48 h. The cells were then treated with glutamine (4 mM) at different time points. Degradation of GLUL was analyzed by immunoblotting. (B) Quantitative results from three independent experiments. (C) GLUL-Myc and V5-Ub were expressed in HEK293T cells. After 24 h, the cells were treated with TD-158 (500 nM) or DMSO for 12 h and then treated with bortezomib (100 nM) or DMSO for 12 h. Whole-cell lysates and proteins immunoprecipitated using Myc magnetic beads were analyzed by immunoblotting for the indicated proteins. (D,E) WSU-DLCL2 (D) and RPMI8226 (E) cells were pre-treated with TD-165 (1 μM) or DMSO for 24 h and, after harvesting, were divided into four groups. Each group was then treated with pomalidomide (1 μM) and DMSO, or pomalidomide (1 μM) and TD-165 (1 μM), for 3 d. Cell viability was measured using CellTiter-Glo (**P < 0.001, *P < 0.01).
