Fig. 4

Mutation of the SUMOylation sites in eIF5A1 modulates its subcellular localization. A U2Os cells were transfected with HA–eIF5A1–WT, HA–eIF5A1–3KR, or HA–eIF5A1–5KR and 36 h after transfection localization of eIF5A was evaluated using immunofluorescence assay with anti-HA antibody. The nuclear to cytoplasmic ratio was analyzed from images using ImageJ analysis software. Data represent the mean and error bars of 50 cells. Statistical analysis was assessed by a Student’s t-test.***P < 0.001. B HEK-293 cells were transfected with HA–eIF5A1–WT, HA–eIF5A1–3KR, or HA–eIF5A–5KR, as indicated. Thirty-six hours after transfection, cells were subjected to subcellular fractionation and the levels of eIF5A1 protein in the nucleus and cytoplasm were evaluated using western blot analysis with anti-HA antibody. C HEK-293 cells treated with DMSO or the SUMOylation inhibitor ML-792 were subjected to subcellular fractionation and then the levels of the eIF5A1 protein in the nucleus and cytoplasm were evaluated using western blot analysis with anti-eIF5A1 antibody