Figure 3.

Individual phosphomutations at Ser-10 or Ser-25 alter ICAP1 nuclear localization. To assess whether single serine mutations alter ICAP1 localization, individual GFP-ICAP1 serine phospho-mimicking or phospho-blocking mutations from group i (A–C) or group iii (D–F) were generated and expressed in CHO cells. A and D, representative images of CHO cells expressing GFP-tagged ICAP1 constructs stained with DAPI (to identify nuclei) 24 h after plating on fibronectin. Bar, 10 μm. B and E, percentage of GFP intensity in the nucleus. Boxes, 25–50th and 50–75th percentile; whiskers, 10–90th percentile; +, mean. Results are from three independent experiments, and the total number of cells examined (N) is indicated for each condition. ****, statistical significance at p ≤ 0.0001 as determined by a one-way ANOVA with Fisher's LSD test with multiple comparisons. C and F, representative immunoblots (against anti-GFP and anti-vinculin) (IB) indicate expression of individual phosphomutants at the expected size. Uncropped immunoblots are shown in Fig. S1 (B and C).