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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 2. — TcBuster transposase rodlet number increases with increasing plasmid DNA doses. HEK-293 cells on coverslips in 6-well plates were transfected by FuGene 6 with 500 ng of transposon (pTcBNeo) and varying doses of pCMV-HA-TcBuster (0 ng, 10 ng, 50 ng, 100 ng, 250 ng or 500 ng), and pUC19 as needed to increase the DNA amount to 1000 ng into HEK-293 cells in six-well dishes. Representative images of cells given 0 ng (A), 50 ng (B), 100 ng (C) or 500 ng (D) of pCMV-HA-TcBuster are shown (DAPI-stained DNA is in blue; HA-TcBuster transposase is stained in red). At each dose, two coverslips were stained and three images per coverslip were taken of random of fields containing at least one rodlet. The average and standard error of the six images taken based on data from blinded counts are plotted (E–G). (E) The number of rodlets per HA-TcBuster positive nuclei is shown for various transposase plasmid doses. (F) The percentage of nuclei with rodlets was calculated at each DNA dose. (G) The number of rodlets as a fraction of all nuclei. Lines of best fit are displayed with the corresponding equation and R² value for each (E–G).