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Fig. 5. FRAP analysis of GFP-MBNL1 mobility in DM1 fibroblasts. DM cells (GM3987, GM3132) with CTG repeats of different lengths (500 and 2000 repeats, respectively) were transfected with GFP-MBNL1. (A) GFP-MBNL1 colocalizes with nuclear foci of endogenously expressed CUG repeat RNA. Arrows indicate the location of the foci. CUG-containing RNA was detected using Cy3-labeled (CAG)5 antisense PNA probe, nuclei were counterstained with DAPI. Bar, 10 µm. (B) Photobleaching of foci and nucleoplasm in DM1 fibroblasts (~2000 CTG repeats) transfected with GFP-MBNL1. Circles denote the photobleached regions in the foci and nucleoplasm. Bar, 10 µM. (C) Recovery curves of photobleached foci and nucleoplasm in DM1 fibroblasts. GM3987 and GM3132 fibroblasts were transfected with GFP-MBNL1. The graph represents data collected for 37 GM3132 fibroblasts and 31 GM3987 fibroblasts over three independent experiments.





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