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. 2015 Sep;21(9):1532–1543. doi: 10.1261/rna.051524.115

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© 2015 Haag et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society

This article, published in RNA, is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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FIGURE 4. — NSUN6 localizes to the cytoplasm and partially colocalizes with pericentriolar matrix and Golgi apparatus marker proteins. (A) HEK293 cells stably expressing NSUN6-HisPrcFLAG were separated into nuclear and cytoplasmic fractions. Nuclear and cytoplasmic protein extracts from equal numbers of cells were separated by SDS-PAGE and analyzed by Western blot. NSUN6-HisPrcFLAG was detected using an anti-FLAG antibody; UTP14A and tubulin were used as nuclear and cytoplasmic markers, respectively. (B,C) HEK293 cells stably expressing GFP-NSUN6 (B) or NSUN6-GFP (C) under the control of a tetracycline inducible promoter were examined by fluorescence microscopy (green in overlay). The pericentriolar matrix (PCM) or Golgi apparatus (red in overlay) was stained by immunofluorescence using anti-γ-tubulin (MTOC), anti-PCM1 (PCM), anti-GM130 (cis-Golgi), or anti-GOPC (trans-Golgi) antibodies. White arrows indicate colocalization of NSUN6 with γ-tubulin (MTOC) and PCM1 (PCM). Scale bar: 20 µm.