Science - USA (2021-12-10)

Zimmerliet al.,Science 374 , eabd9776 (2021) 10 December 2021 8 of 15

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ABcontrol osmotic shock

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Fig. 7. NE tension is reduced during ED and OS and leads to NPC constriction.
(A) Maximum projections of Nup60-mCherry channel from live-cell imaging
confocal stacks of NLS-GFP-NLS and Nup60-mCherry–expressing cells used for
nuclear shape and volume analysis. Scale bar, 5mm. (B) Same as (A) but for
cells expressing ubiquitous sfGFP and Nup60-mCherry under OS conditions.
(CandD) Quantification of nuclear volume based on 3D reconstructions of Nup60-
mCherry signal corresponding to (A) and (B). Data are from three experiments,
and triangles indicate means of individual replicates (means ± SDs;n= 300 nuclei
per condition under ED andn= 300 nuclei per condition under OS conditions).
One-way ANOVA and Dunnett’s multiple comparison test; ***P< 0.001;

**P< 0.01. (E) Number of NPCs per estimated NE surface area under control,

ED, OS, and OS recovery conditions (see Materials and methods). Data are from

one or more experiments (means ± SDs;n= 71 tomograms under control,

n= 72 tomograms under ED,n= 33 tomograms under OS, andn= 53 tomograms

under OS recovery conditions). One-way ANOVA andŠidák’s multiple comparison

test; ns indicates not significant,P> 0.05. (FandG) Deconvolved slice through

representative tomograms under control (F) and OS conditions (G). N marks

the nucleus, and arrowheads mark NPCs. Scale bars, 200 nm. (H) Mean NPC

diameter (see also Fig. 6A) plotted against mean nuclear volume [same as (C) and

(D)] under and control and 60 min after ED and OS conditions. Linear regression

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