Removal of the transmembrane segments reduces effects of NETs in chromosome repositioning. Soluble fragments of NETs from their largest predicted nucleoplasmic segment were fused to an NLS and stably expressed in HT1080 cells to determine if they could influence chromosome repositioning. (a) Schematic diagram of each NET highlighting the soluble fragment in blue. The dark grey boxes indicate predicted transmembrane spans. (b) Images showing nucleoplasmic targeting of the GFP fusion constructs. (c) Cells expressing soluble fragments encoding the principal nucleoplasmic regions of NETs did not recapitulate the strong effects in repositioning observed with full-length NETs (Figure 5b). Error bars indicate standard deviation between the means of individual experiments. *P < 0.05 comparing the position of the chromosome in the NET-transfected cells to the NLS-GFP transfected control using KS tests. None of the soluble NET fragments yielded higher stringency P-values < 0.01. Statistics are given in Additional file 1.
Zuleger et al. Genome Biology 2013 14:R14 doi:10.1186/gb-2013-14-2-r14