Interfering with RanGTP disrupts CAS nuclear retention. (A) Immunostaining of endoKapβ1 in cells expressing mCherry-CAS in the absence and presence of SA and DG. Scale bar, 5 μm. (B) The combined use of SA and DG leads to a marked reduction of mCherry-CAS in the nucleus. (C) Immunostaining of endoRan and endoCAS in the absence and presence of SA and DG. Scale bar, 5 μm. (D and E) (D) EndoCAS and (E) endoRan are reduced in the nucleus when SA and DG are present. (F) Immunostaining of endoCAS and endoKapβ1 in cells expressing either mCherry-Ran or its non-hydrolyzable mutant mCherry-RanQ69L. Scale bar, 5 μm. (G) Scatter plot showing a correlation between the N/C ratios of endoCAS and mCherry-Ran. The N/C ratios of endoCAS and mCherry-RanQ69L are not correlated. Each data point is obtained from an individual cell. Dashed line indicates N/C = 1. Box plots denote the median, first, and third quartiles. The whiskers represent the minimum and maximum values. Median values are indicated next to the box plots. N = 3. The data were tested for normality using a built-in function of GraphPad Prism software. P values were obtained using an unpaired two-tailed t test.