Figure 4.

A DEK-GFP fusion protein is retained on mitotic chromosomes and associates with lagging chromosomes. (A) Schematic of the DEK-GFP fusion protein expressed from a pLentiLox vector backbone. GFP is fused to the C-terminus of the 375 amino acid protein DEK. (B) NIKS were transduced with DEK-GFP or empty control GFP vector and were sorted for GFP expression. DEK-GFP expression was confirmed by western blot analysis using a DEK antibody (BD Biosystems). (C) Quantification of interphase and mitotic cells with chromatin-associated DEK-GFP compared to control GFP protein using a GFP antibody for IF. (D) Mitotic defects were commonly observed in the DEK-GFP expressing NIKS by live cell imaging. Pictures were taken of the same cell in metaphase, anaphase and telophase. DEK-GFP remains bound to the DNA throughout mitosis and is detected on an anaphase bridge that results in a nuclear bud. Arrows point to improperly segregating chromosome(s).