Fig. 5.

SAF-A mutants that release C₀T-1 RNA affect cytological chromatin packaging. For all images: color channels are separated in black and white. Scale bars 5 μm. Cell types: normal human fibroblasts (Tig-1) & mouse/human hybrid cells with human chromosome 4 (Hybrid). A C280-GFP expression causes C₀T-1 RNA to release and DAPI to condense compared to un-transfected neighboring cell. B–C Linescan of C280 expressing cell (B) and untransfected neighboring cell (C) showing larger variation in DAPI intensity measurements for C280 positive cell. C₀T-1 RNA localizes with C280 and not with DAPI. Linescan paths indicated in A). D–F The human Chr4 territory (identified by human C₀T-1 DNA) is larger in control hybrid cells (D) and condenses in hybrid cells expressing the G29A mutant (E), as quantified in (F). F The total volume of the human chromosome territory decreases by 38% in G29A expressing cells (P = 0.0002, n = 11). Error bars, standard deviation of the mean. Z-stacks deconvolved and voxel volume measured using Huygens software