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Figure 3 | Molecular Cancer

Figure 3

From: Nuclear export regulation of COP1 by 14-3-3σ in response to DNA damage

Figure 3

14-3-3σ binds Cop1 and induces COP1 nuclear export via its NES. (A) Identification of 14-3-3σ domain that interactions with COP1. 293T cells were cotransfected with Myc-COP1 and Flag-14-3-3σ wild type (WT) (aa 1-248), N-terminal (aa 1-161) or C-terminal (aa 153-248). Before harvesting, cells were treated with proteasome inhibitors. Lysates were immunoprecipitated with anti-Myc and immunoblotted with anti-Flag. Specific interactions of 14-3-3σ domains with COP1 are indicated. Asterisks indicate IgG light chain. WCL, whole cell lysate. (B) 14-3-3σ nuclear export signal (NES) mutant is unable to mediate cytoplasmic accumulation of COP1. U2OS cells were cotransfected as indicated with GFP-COP1, Flag-14-3-3σ wild type (WT), or RFP-14-3-3σ NES (I205A, L208A) mutant. Live-cell images of GFP-COP1 localization were captured using a fluorescence microscope. The bar graph shows quantification of the nuclear localization of COP1. Error bars represent 95% confidence intervals. Scale bar, 10 μm.

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