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. 2014 Oct 17;29(1):208–215. doi: 10.1096/fj.14-259663

Figure 1.

Figure 1.

MLN4924 deneddylates Cul-2 and stabilizes HIF-1α in HeLa cells. A) Western blotting of HeLa cells treated with vehicle control (veh) or 330 nM MLN4924 for 1 h or increasing doses of MLN4924 indicates deneddylation of Cul-2 and stabilization of HIF-1α in the nuclear fraction. B) Densitometry of the ratio of neddylated:unneddylated Cul-2 protein in HeLa cells treated with vehicle control or 330 nM MLN4924 for 1 h (n = 3 independent experiments). *P < 0.01 C) Western blotting of HeLa cells treated with vehicle control (veh) or increasing doses of MLN4924 for 1 h indicates deneddylation of Cul-2 and stabilization of HIF-1α in the nuclear fraction. D) Western blotting of HeLa cells with 330 nM MLN4924 over a time course demonstrates a loss of NEDD8 conjugated to cullin proteins at T = 5, and a stabilization of HIF-1α and HIF-1α-OH at T = 15 in the nuclear fraction. In all cases, n = 3 independent experiments.