Figure 4
From: The COP1 E3-ligase interacts with FIP200, a key regulator of mammalian autophagy
Ectopic expression of COP1 reduced a form of FIP200 protein. (A) NIH3T3 cells were transfected with expression vectors encoding HA-GFP and HA-GFP-fused COP1. After selection, cells were treated or non-treated with MG132, harvested, and analyzed by Western blotting using antibodies against COP1, FIP200, ULK1, Atg13, Atg101, and γ-tubulin. (B) NIH3T3 cells were transfected with expression vectors encoding HA-GFP and HA-GFP-fused COP1 (wild type and SA mutant). Cell lysates isolated from transfectants after selection were immunoprecipitated with normal rabbit serum (Control) and rabbit antibody to an HA-epitope (α-HA). Immune complex was analyzed by Western blotting using antibody against COP1. (C) Cells in panel B were treated with UV then incubated in complete medium for 0, 8, and 16 hours. Cells were harvested, and analyzed by Western blotting using antibodies against p53, FIP200, and γ-tubulin.