Fig. 3.

Cullin neddylation is not required for emetine-induced HIF-2α degradation. 786-O cells were transfected with either control siRNA or NEDD8 siRNA. A, to confirm NEDD8 silencing, NEDD8 mRNA was analyzed by quantitative RT-PCR 72 h after transfection. Statistical significance was calculated using SigmaPlot 9.0 (*, p < 0.05) (Systat Software Inc., San Jose, CA). Each bar represents the mean ± S.D. (n = 3). B, 786-O cells were transfected with control or NEDD8 siRNA as in A, and after 72 h, the cells were treated with 0.3 μM emetine for an additional 4 h. HIF-2α protein level was assessed in nuclear lysates by Western blotting. CREB expression is shown as a loading control.