Fig. 4

SUMOylation of hSSB1 stabilizes the protein. a–f The indicated cell lines were treated with the indicated concentrations of the SUMOylation inhibitor 2-D08, and 24 h later, the cells were subjected to western blotting or real-time PCR. g HEK293T cells cotransfected with FLAG-hSSB1 and HA-ub were treated 24 h later with the indicated concentrations of 2-D08 for 24 h, and then the cells were lysed and analyzed by western blotting or IP using the anti-FLAG antibody followed by western blotting. h HEK293T cells transfected with the indicated plasmids were treated 24 h later with 20 μg/ml cycloheximide (CHX) for the indicated times, and then the cells were lysed and analyzed by western blotting. i Quantification of the protein half-lives based on h. The results shown are averages of three independent experiments. Bars indicate the SEM. j HEK293T cells transfected with the indicated plasmids for 48 h were lysed and analyzed as in g