Fig. 4. Gomafu RNA forms a novel nuclear domain. (A-K) The subnuclear localization of Gomafu RNA was examined by fluorescent in situ hybridization and compared with various nuclear domain markers in E14 retinal cells (A-F) and in HeLa cells transfected with a vector expressing Gomafu cDNA (G-K). In D, cells were pre-treated with actinomycin D (ACTD). Note that Gomafu RNA signals (green) did not coincide with immunolabeled signals for SC35 (C,D,G), fibrillarin (Fbl, E,I), PML (F,J), PSP1 (H), and coilin (K), which are shown in magenta. (L-N) Double in situ hybridization of E14 retinal cells for Gomafu RNA (green) and other mRNA-like noncoding RNA Evf-1 (L), Xist (M) and Air (N), which were shown in magenta. All the images are single optical sections obtained using a laser-scanning confocal microscope. Bars, 2 µm.