Pericentromeric targeting of the mutant IKAROS. NIH3T3 cells were transfected with a hemagglutinin (HA)-tagged WT or mutant expression vector alone (A) or together (B) with Flag-tagged WT IKAROS. Cells were labeled with anti-HA or together with anti-Flag antibodies, followed by Alexa Fluor 488–conjugated (green) and Alexa Fluor 568–conjugated (red) secondary antibodies. Cells were visualized using a ZOE fluorescence microscope (original magnification, ×175). Data are representative of 3 independent experiments. HI and DN indicate DNA binding–defective haploinsufficient mutation and DN mutation, respectively. Scale bars, 25 μm.