Fig. 8: Recapitulation of RanGTPase-controlled nuclear import with the FG phase assembled by the perfectly repetitive variant.
From: Recapitulation of selective nuclear import and export with a perfectly repeated 12mer GLFG peptide
a The FG phase of prf.GLFG52x12 was initially loaded with hsImpβ·hsIBB-EGFP complex. At time = 0 s, a RanQ69L fragment fused to MBP-mCherry was added, which triggered the unloading of IBB-EGFP from Impβ and thus an efflux from the FG phase. Fluorescence signals of GFP and mCherry were recorded over time. The ratios of GFP fluorescence inside:outside an FG particle of 7 μm radius (marked with a white arrow) were quantified. Note that some Ran fusion was recruited to the FG phase by Impβ but only arrested at the rim of the FG particles, as expected from the FG-phobic effect of the MBP-mCherry group. This was meant to ensure that the RanGTP-reaction occurs only at the particles’ surface. b Time course of IBB-EGFP signal inside the FG phase. Blue colored: GFP signal inside an FG particle of 7 μm radius (marked with a white arrow in (a)), normalized to % of the initial signal. Gray: a control set without RanGTP addition. Orange: best-fit to a single exponential decay function: \(f\left(t\right)=A{e}^{-{kt}}+B\), where t is the time, A = 110, B = 0, k = 4.4 × 10−3. c The GFP signals inside FG particles with different radii were fitted to the single exponential decay function to obtain the respective time constants k. Source data are provided as a Source Data file. d k obtained in the experiment described above was plotted against 1/r2, where r is the FG particle’s radius. e Illustration of the above processes.
