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. 2019 Oct 16;39(1):e101689. doi: 10.15252/embj.2019101689

Figure 1. Quantitative FLIM/FRET imaging reveals pleiotropic effects of RanT24N on chromosomal gradients and importin β‐cargo binding.

Figure 1

  • A
    Schematics of Rango‐3 sensor.
  • B
    Representative images of importin β cargo gradients detected by Rango‐3 FLIM/FRET imaging. Scale bars, 20μm.
  • C
    Line scans of Rango‐3 FRET efficiency (E) in 10‐μm‐wide areas centered at the dashed lines in (B).
  • D–F
    Quantification of the Rango‐3 E gradients (top panels) and average cellular Rango‐3 E (bottom) in involving untreated oocytes (controls) or oocytes injected with RanT24N mRNA, importin β(71–876) protein or treated with 20 μM IPZ. Data for each panel are from at least 2 separate experiments. Means ± SDs, t‐test, oocyte numbers indicated in brackets.
  • G
    Nonlinear single‐exponential regression was used to fit the average of radial Rango‐3 τdonor line scans in control oocyte (Fig 1D) with the one‐phase decay model. The dashed line corresponds to the average of spindle pole distance from chromosomes in the same cell.
  • H
    Immunoblotting in biochemical pulldowns with recombinant Ran proteins added to human DLD1 cell lysates. A representative of at least five repeats.