Modification efficiencies obtained testing different concentrations of mimosine, thymidine, vinblastine and SDFs with different superimposed methylation patterns on D1 cells.

<p>A) For each drug the concentration that gives the highest percentage of synchronized cells and the lowest cell death (highlighted in grey) was selected. B) Correction efficiencies after transfection in different cell cycle phases. A SDF homologous to mutated eGFP sequence was used as control (CTR). Gene modification efficiency was enhanced when cells are synchronized in G₂/M phase (*p = 0.0001 respect to CTR and +p = 0.0001 respect to unsynchronized cells, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030851#pone.0030851.s005" target="_blank">Fig. S5</a>). C) Differently <i>in vitro</i> methylated SDFs were tested to assess methylation involvement in gene modification efficiency. SDF-PCR-WT gave the highest efficiency of modification (*p resulted to be significant when compared to all treatments; specifically p = 0.002 respect to <i>Dam+</i>, p = 0.01 respect to <i>SssI</i>+, p = 0.008 respect to <i>Dam+/SssI+</i>, and p = 0.009 respect to SDF-DIG-WT). For representative FACS dot plots see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030851#pone.0030851.s005" target="_blank">Fig. S5</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030851#pone.0030851.s006" target="_blank">S6</a>.