Skip to main content
. 2019 Jul 1;116(29):14606–14613. doi: 10.1073/pnas.1903764116

Fig. 1.

Fig. 1.

Description of the NuRIM method. (A) State-of-the-art electron density map of the yeast NPC adapted from the work of Kim et al. (5) introduces salient features of the NPC including a central channel permeated by a central “transporter,” framed by the NPC core. UCSF Chimera software was used to display a density map at the recommended threshold of 0.015 (64). (B) Bright-field image of S. cerevisiae cells. (C) The luminal dsRed-HDEL reference is used for NE tracing. (D) Using automated image analysis, NEs are precisely outlined and their center is determined (cross-hair). (E) Nup-yEGFP channel allows quantifying Nup abundance and radial shift relative to the dsRed-HDEL reference channel. (F) Dilated NE traces restrict analysis to informative area only. (G–I) Lines from the nuclear center intersect the NE. Intensity profiles along these lines are then fitted with Gaussian functions, allowing the measurement of the radial shift of a given Nup, for example, Nup159-yEGFP (green) against the dsRed-HDEL reference profile along the same ray (red). Averaging thousands of such differential measurements delivers structural information on NPC architecture with nanoscale accuracy (SI Appendix). (Scale bars, 1 μm.)