Fig. 3

SUMOylation of SET at K68 induces its cytoplasmic retention. a HEK-293 T cells were co-transfected with Flag-SET-WT, Flag-SET-K68R, His-SUMO-1 and UBC9 plasmids for 48 h. Cells were lysed and probed for SET via western blotting analysis. b Quantification of the blots described in (a) was performed using ImageJ software. c and d Cytosolic and nuclear fractions were prepared in cells co-transfected with Flag-SET-WT or Flag-SET-K68R, His-SUMO-1 and UBC9 for 48 h. Levels of SET in the cytoplasm and nuclear were determined by western blotting with anti-SET antibodies. Relative purity of the fractions was confirmed by sequential probing for the cytoplasmic marker Raf-1 and the nuclear marker LaminB1. e and f Quantification of the blots in C-D. **P < 0.01