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. 2016 Nov 29;45(1):353–366. doi: 10.1093/nar/gkw1115

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Published by Oxford University Press on behalf of Nucleic Acids Research 2016. This work is written by (a) US Government employee(s) and is in the public domain in the US.

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Figure 3. — Colocalization within rodlets following delayed transfection of differently tagged transposase constructs. (A) To test the activity of the Flag-tagged TcBuster transposase, HEK-293 cells were transfected with the transposase plasmid indicated and the pTcBNeo neomycin-resistance transposon plasmid at a 1:9 ratio, n = 3 per group. After 48 h, the cells were split into selection media and colonies were counted after two weeks of selection. To examine the dynamics of rodlet formation, HEK-293 cells grown on coverslips in six-well plates were transfected by FuGene 6 with pCMV-TcBuster (500 ng) and pCMV-HA-TcBuster (500 ng) then 6 h later with pCMV-TcBuster (500 ng) and pCMV-Flag-TcBuster (500 ng) (B) or vice versa (C). The following day, the cells were double-stained for Flag-tagged TcBuster transposase (green) and HA-tagged TcBuster transposase (red). Cells transfected with one of the transfections produced red or green rodlets only, whereas cells transfected with both produced only uniformly yellow rodlets as shown in the representative images including a 4× magnification of a nuclei with rodlets costaining for Flag-TcBuster and HA-TcBuster, inset.