FIGURE 3.
ICAP1FL is more nuclear than ICAP1PTB. A, CHO cells overexpressing GFP-tagged ICAP1 constructs were plated on fibronectin, fixed 24 h later, and stained with DAPI to identify nuclei. Representative images are shown; bar, 10 μm. B, percentage of GFP intensity in the nucleus compared with the integrated GFP intensity of the entire cell was calculated using CellProfiler version 2.0. Boxes, 25th through 50th and 50th through 75th percentile; whiskers, 5th through 95th percentile (n = 97–130 cells from 5 independent experiments). *, p ≤ 0.0001 as determined by a one-way ANOVA with Tukey's correction for multiple tests. C, representative fractionation of CHO cells overexpressing GFP-tagged ICAP1 constructs. C, 28% of the cytoplasmic fraction; N, 80% of the nuclear fraction. Carbonyl reductase (CBR1) and histone deacetylase (HDAC1) represent quality controls for cytoplasmic and nuclear fractions, respectively. D, quantification of cell fractionation data, where the percentage nuclear = total nuclear/(total nuclear N + total cytoplasmic) × 100 (bar, mean percentage nuclear value; n = 10). *, p ≤ 0.001 as determined by a one-way ANOVA with Tukey's correction for multiple tests.