Figure 8.

NANOG localization during the cell cycle. (a,c) Analysis of NANOG during the cell cycle by double indirect immunofluorescence. NANOG (green; #4903, Cell Signaling Technology (CST)) colocalized with one centrosome (pericentrin, PCNT; red) only during interphase (a) but with both centrosomes (pericentrin, PCNT; red) during mitosis (c). The nuclei were counterstained with Hoechst 33342 (blue). For each image, regions of interest are indicated by the dashed and solid boxes, and the close-ups are provided on the right and below, respectively. Scale bars, 5 μm. (b) Schematic visualization of the centrosome duplication cycle: The centrosome is copied only once per cell cycle following cytokinesis. The centrosome duplication cycle results in two daughter cells with different ages of centrosomes: one that keeps the mother centrosome with the old mother centriole and the other cell that inherits the daughter centrosome, which comprises the young mother centriole [51,52]. Distal appendages (light blue) remain on the mother centriole during the whole cell cycle and appear during mitosis or shortly after mitosis on the older centriole of the daughter centrosome [43,53]. After mitosis, the newly formed mother centriole assembles subdistal appendages (blue) [43,53].