Fig. 7: Mobility of NTRs inside FG phases assembled from Pro-containing perfect repeats.
From: Recapitulation of selective nuclear import and export with a perfectly repeated 12mer GLFG peptide
a, b FG particles (unlabeled) assembled from both prf.GLFG52x12 versions were preloaded with Alexa488-labeled NTF2, bleached, and FRAP was recorded over time. Experiments were repeated with multiple FG particles (n = 5). Representative images (a) and recovery curves (b) are shown. See Supplementary Figs. 5 and 6 for the complete dataset and other FRAP datasets for GFP-NTR derivatives. Source data are provided as a Source Data file. c FG particles spike with Alexa647 (covalently)-labeled FG domain molecules (FG-tracers) were immobilized on glass, preloaded with Alexa488-labeled NTF2, washed multiple times with fresh buffer (“Methods”) while the fluorescence signals were recorded. Orange numbers: % Alexa488 signal inside FG phase relative to the initial signal. For both Mac98A and prf.GLFG52x12 (GLEBS-free), signals of NTF2 within FG particles dropped to ≈20% of the original after seven rounds of washing. The majority of FG particles remained immobilized after washing, indicating their stickiness to the support.
