FIGURE 3:
SENP1 and SENP2 are targeted to overlapping and distinct mitotic structures through their N-terminal domains. Constructs coding for GFP-tagged SENP1, SENP2, or SENP1/2 chimeras were transfected into HeLa cells. Cells expressing SENP1 or SENP1N2cat were permeabilized, fixed, and stained, where cells expressing SENP2 or SENP2N1cat cells were fixed, permeabilized, and stained. Cells were colabeled with (A) CREST anti-centromere antibodies (asterisks highlight centrosome staining, and arrowheads indicate spindle microtubule staining), (B) antibodies specific for the inner-centromere marker INCENP (arrowheads indicate centromeres flanked by kinetochore-associated SENP1 and SENP2 signals), and (C) antibodies specific for the outer-kinetochore marker Hec1. DNA was labeled with DAPI. Cells were imaged by immunofluorescence microscopy. Bar, 10 μm.