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. 2011 Jun 3;12(7):735–742. doi: 10.1038/embor.2011.86

Figure 5.

Figure 5

Expression of expanded CTG repeats results in mislocalization of SHARP in COS7 and HEK293 cells. (A) The constructs DMPK 11–15(CTG)5 and DMPK 11–15(CTG)300 were expressed in COS7 cells. Endogenous SHARP (green, a,e) and CUG foci (red, b,f) were visualized 48 h post-transfection. Nuclei were stained with DAPI (c,g). Merged images (d,h) demonstrate that SHARP localizes primarily within the nucleus of cells expressing the DMPK 11–15(CTG)5 and predominantly in the cytoplasm of cells expressing expanded (CTG)300 tracts. (B) Similar results were obtained in HEK293 cells transfected with DMPK 13–15(CTG)5 and DMPK 13–15(CTG)700. Endogenous SHARP is shown in green (a,e,i), CUG foci are shown in red (b,f,j), nuclei stained with DAPI (c,g,k) and merged images (d,h,l) are shown. Scale bar, 20 μM. CMV, cytomegalovirus; DAPI, 4,6-diamidino-2-phenylindole; DM1, myotonic dystrophy; SHARP, SMART/HDAC1 associated repressor protein.