Figure 3.

Impaired SV40 NLS-GFP and hnRNPA1 NLS-GFP Import into the Nucleus in Affected Individuals

Representative immunofluorescent images showing less GFP (green) transport into the nuclei of affected individuals’ fibroblasts compared to the GFP transport into the nuclei of control fibroblasts. Disruption of NUP188 in affected individuals inhibited the “classic” importinα-importinβ1 nuclear import pathway (SV40 NLS) and the transportin-1 pathway (hnRNPA1 NLS) but not the rate of diffusion (no NLS). DAPI (blue) was used to define the boundaries of the nucleus, and wheat-germ agglutinin (not shown) was used to define the cell boundaries for quantification of the ratio of nuclear GFP fluorescent intensity to cytoplasmic GFP fluorescent intensity. Error bars represent SEM. The scale bar represents 20 μm. The experiment was performed in triplicate with a combined n = 47–70 per condition, representative images shown. ^∗∗p < 0.01, ^∗∗∗∗p < 0.0001 by two-tailed unpaired Student’s t test.