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Description: Figure 3. Nuclear Localization of snolncRNAs (A) Each PWS region sno-lncRNAexhibits strong nuclear accumulation in PA1 cells. RNA ISH (green) was performed with individual probes recognizing the internal sequence of each snolncRNA, or with pooled probes recognizing all PWS regionsno-lncRNAs (bottom right). Representative images are shown. DAPI is in blue and the white scale bar in all images denotes 10mm. (B) Sno-lncRNAs do not accumulate in either nucleoli or CBs. Costaining of sno-lncRNAs (green) and marker proteins for nucleoli (Nucleolin, red) and CBs (Coilin, red), respectively in H9 cells. Representative images are shown. (C) Sno-lncRNAs accumulate at a single chromosomal locus. Double FISH ofsno-lncRNAs (green) and its adjacent DNA region (red) in both PA1 and H9 cells. A single Z stack of representative images acquired with an Olympus IX70 DeltaVision Deconvolution System microscope is shown. (D) Total RNA from PA1 cells was separated into cytoplasmic (not shown), nuclear soluble, and nuclear insoluble fractions. Bar plots represent relative abundance of RNAs in the nuclear soluble and insoluble fractions as measured by RT-qPCR. Data are collected from Figure 3.
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