Aurora C signals appeared as substantial vibrant nuclear staining corresponding towards the heterochromatic chromocenters usually observed at the nuclear periphery. These chromocenters represented clustered centromere heterochromatic areas of chromosomes. Fig. 3A also demonstrates that Aurora C was colocalized with Aurora B and INCENP at the chromocenters in diplotene spermatocytes. Interestingly, the CENP H antibody recognized sister kinetochores, which appeared as pairs of dots found on major on the Aurora C signals. The physical appearance of Aurora GS-1101 distributor B and INCENP in diplotene spermatocytes agrees having a earlier report. For the duration of metaphase I, Aurora C was colocalized with AuroraB and INCENP mainly while in the centromeric regions. Interestingly, nearly all of the Aurora C labeling was detected beneath the kinetochore CENP H signals, whilst some degree of overlap was observed. Thus, Aurora C is more than likely found involving CENP H and the heterochromatin. At the onset of anaphase, Aurora C, like Aurora B, was transferred in the centromeres to the spindle midzone and was at some point concentrated in the midbody.

To examine the localization of Aurora C in the course of the diakinesis to metaphase I transition in far more detail, immunofluorescence staining Cellular differentiation of chromosome spreads of meiotic cells was carried out. Remarkably, a considerable quantity of AuroraC signal was detected along the chromosomal axes, which covered each the regions with the centromere and also the chromosome arms for the duration of diakinesis. Intense Aurora C signals were usually observed during the arm areas proximal towards the centromeres. At the MI stage, nevertheless, most Aurora C signals were detected at the centromeres. Very similar effects have been also observed for Aurora B kinase. By evaluating Aurora C signals between the diakinesis and MI phases, it is reasonable to speculate that Aurora C gradually dissociates from the arms and accumulates on the centromeres during the diakinesis to MI transition.

Due to the fact really couple of cells are present at this transition CTEP stage in the course of regular meiotic division, we treated pachytene spermatocytes with okadaic acid, a protein phosphatase inhibitor. It has been reported that OA can induce a speedy and premature G2/M transition that is accompanied with the disassembly of SCs. Just after OA remedy, discontinuous signals of Aurora C dotted along the chromosome arms were plainly noticeable in some OAFig treated cells, more than likely representing the diakinesis to MI transition. While in many others, Aurora C signals have been prominently identified at the centromeres of MI spermatocytes. Together, our results recommend that Aurora C is localized along the chromosome arms and centromere regions, when its arm association is gradually misplaced through the diakinesis to meiosis I transition.