1. NPM1 drives poly-GR into the nucleolus of primary neurons.
NPM1 drives poly-GR into the nucleolus of primary neurons. Primary rat neurons (DIV7 + 7) were cotransduced with RFP-tagged nucleolar interactors NPM1 and NOP56 with GFP, GFP-(GR)149, or (PR)175-GFP. (A–C) Single focal planes obtained by confocal microscopy are shown. DAPI was used as nuclear marker and the scale bar depicts 20 μm. RFP was used as negative control. Left columns show GFP signal, middle columns show RFP-tagged proteins, and right columns show merge of GFP, RFP-tagged proteins, and nuclear DAPI staining (blue). RFP-NPM1 and RFP-NOP56 are co-localizing with poly-PR. Note that NPM1 expression recruits poly-GR into the nucleolus (red arrow). Poly-PR–expressing neurons show altered NOP56 nucleolar staining (single channel shown in zoom). (D) Fraction of cells with poly-GR localized to the nucleolus in NPM1– and NOP56–expressing neurons compared with the RFP control (RFP, n = 9; RFP-NPM1, n = 11; and RFP-NOP56, n = 10 images (40× objective) from three independent experiments, mean ± SEM, exact P-values: RFP versus RFP-NPM1, P = and RFP versus RFP-NOP56, P = in one-way ANOVA with Dunnett’s posttest).
Hannelore Hartmann et al. LSA 2018;1:e
© 2018 Hartmann et al.