Fig. 2. The C-terminal end of NIP2 is critical for centriolar localization. (A) U2OS cells were placed on ice for 30 minutes to disrupt microtubules or treated with 1 µg/ml taxol for 2 hours to stabilize them, and were immunostained with antibodies against NIP2 and β-tubulin. Arrows indicate the endogenous NIP2 staining at the centrosomes. Bar, 5 µm. (B) Several NIP2-truncated mutants with the GFP tags were expressed in HeLa cells. The β-tubulin was detected as a control. (C) HeLa cells were transfected with the NIP2-truncated mutant genes and immunostained with antibodies against GFP (green) and -tubulin (red). Arrows indicate the centrosomes. Bar, 10 µm. (D) Myc-tagged NIP2 truncated mutant proteins (Myc-NIP2, Myc-NIP2^445-903, and Myc-NIP2^1-523) were co-expressed with GFP-centrin 2 (green) in HeLa cells and detected with the Myc antibody (red). The insets are magnified views of the centrosomes.