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. 2016 Feb 8;7:10624. doi: 10.1038/ncomms10624

Figure 2. Light-dependent inhibition of endogenous nuclear export.

Figure 2

(a) Schematic of blue light-dependent CRM1 sequestration by LEXY fused to H2B-GFP. (b) Schematic of constructs encoding H2B-GFP fused to LEXY and a NLS-mCherry-NES reporter. (c) Fluorescence images and (d) corresponding quantification of the relative nuclear mCherry fluorescence of cells co-transfected with the indicated constructs. Cells were incubated in the dark for 3 min followed by blue light irradiation for 20 min and dark recovery for 20 min. AsLOV2-NES 27, a particularly strong NES variant (Supplementary Fig. 2b); wt, wild type. Data represent box plots and individual data points, n=34 cells for H2B-GFP-AsLOV2, n=42 cells for H2B-GFP-LEXY and n=28 cells for H2B-GFP-AsLOV2-NES27, all from 3 independent experiments; ***P=9.10 × 10−10 (wt AsLOV2 versus LEXY) and 3.45 × 10−5 (LEXY versus AsLOV2-NES27) by Mann–Whitney test. (c) Scale bar, 20 μm. (e) Scatter plot of the relative nuclear mCherry fluorescence versus nuclear EGFP fluorescence for the indicated samples in d after blue light irradiation (23 min time point).