RNA Foci Visualization via LNA Probe

A 5′ TYE-563-labelled LNA (16-mer fluorescent)-incorporated DNA probe was used against the sense (Exiqon, Inc.; batch number 607323) and the antisense RNA hexanucleotide repeat (Exiqon, Inc.; batch number 610331). Slides were prepared and RNA foci were visualised as described previously [3 (link)] using a Leica SP5 confocal microscope system with a ×63/1.4 oil immersion objective lens. Briefly prehybridisation was followed by overnight hybridization at 66 °C in a humid atmosphere. A single wash at room temperature with 2 × SSC/0.1 % Tween-20 preceded three washes at 65 °C with 0.1 × SSC. Slides were then mounted in DAPI Vectashield or processed further for dual staining of RNA and protein.