Fig. 10
From: 14-3-3 proteins regulate cullin 7-mediated Eag1 degradation
The CNBHD and PAS domain are essential for Eag1 regulation by difopein. A Structural topology for Eag1, hErg, and various Eag1 chimeric constructs. Chimera A: Eag1 containing hErg C-linker. Chimera B: Eag1 containing hErg CNBHD. Chimera C: Eag1 containing hErg post-CNBHD region. Chimera N: Eag1 containing the complete hErg N-terminal region. Chimera P: Eag1 containing hErg PAS domain. Chimera O: Eag1 containing hErg N-linker. B, C Replacement with hErg CNBHD (chimera B), PAS domain (chimeras N and P), or N-linker (chimeras N and O) abolishes the effect of difopein on Eag1 protein level. (Left panels) Representative immunoblots. (Right panels) Quantification of relative Eag1 protein levels. Myc-tagged Eag1 wild-type (WT) and chimeric constructs were co-transfected with YFP vector, YFP-R18 mutant or YFP-difopein in HEK293T cells. Protein densities were standardized as the ratio of Eag1 signals to the cognate β-actin signals, followed by normalization with respect to the YFP vector control (*, P < 0.05; n = 3–6)