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. 2015 Jul 28;5:12521. doi: 10.1038/srep12521

Figure 2. FL expHTT RNA forms nuclear foci in the SK-N-MC neuroblastoma cell line.

Figure 2

(A) Schematic representation of FL-HTTQn plasmids. (B) Cells from the SK-N-MC neuroblastoma line were transfected with pcDNA3.1, FL-HTTQ23 or FL-HTTQ82 plasmids, and, 48 hours post-transfection, FISH was performed to detect CAG RNA foci using a 5′ Texas Red-labeled CUG20 riboprobe (red, arrows). Hoechst was used as a nuclear marker. Green fluorescence was imaged as a background control. FL HTT RNA with 23 CAG repeats (FL-HTTQ23) formed rare foci, whereas RNA foci were frequently formed by FL HTT RNA with 82 repeats (FL-HTTQ82). (C) SK-N-MC cells were transfected with FL-HTTQ82 plasmid, treated with DNase or RNase, and then subjected to FISH. Persistence of RNA foci after DNase treatment and absence of the foci following RNase treatment (arrows) confirms that the foci contain RNA. Scale bar, 10 μm.