A, A549 cells were treated with 1 μg/mL doxorubicin (DOX) for the indicated hours. Lysates were analyzed by immunoblotting with indicated antiboies (Left). A549 cells were treated with 1 μg/mL doxorubicin (DOX) for the indicated hours and treated with or without MG132 for 3 hr. Lysates were immunoblotted with the indicated antibodies (Right).
B, Indicated cells were treated with 1 μg/mL doxorubicin (DOX) for the indicated hours. Cell lysates were immunoblotted with the indicated antibodies. The protein levels of COP1 in response to DNA damage at the different time points were determined (Right).
C, HCT116 cells were knocked down with two specific 14-3-3σ shRNA (1 & 2) or control Luciferase shRNA (C). Lysates were immunoblotted with anti-COP1, and anti-14-3-3σ (Left). 293T cells were transfected with the indicated expression vectors. Equal amounts of cell lysates were immunoblotted with the indicated antibodies (Right).
D, 293T cells were transfected with the indicated plasmids. Cells were treated with or without proteasome inhibitor MG132 before lysates were collected. Lysates were immunoblotted with the indicated antibodies (Left). Indicated cells were treated with cycloheximide (CHX) for the indicated hours. Cell lysates were immunoblotted with indicated antibodies. The protein levels of COP1 over time are shown (Right).