Figure 3.
Figure 3. Analysis of inner centromeric H2B S6ph localization. (A) Synchronized RPE-1 cells were used to produce chromosome spreads, which were stained with antibodies specific for H2B S6ph (red) and CENP-A (green). Bar, 1 µm. The images were quantitatively analyzed with ImageJ for the distribution of H2B S6ph and CENP-A along the centromere cross section, as shown by the dotted line (top). The maximum of fluorescence intensity was set as 1, and shaded areas indicate standard deviations (n = 8 cells/368 centromere pairs). (B) The experiment was done as in Fig. 2 C, with the difference that chromosomes were stained for H2B S6ph and the cohesin component RAD21. Representative examples are shown; the boxed area is displayed in the bottom panel in 4× magnification. Bars: 5 µm (main); 1 µm (magnification). (C) Schematic representation of the chromosomal localization of H2B S6ph on mitotic chromatids.

Analysis of inner centromeric H2B S6ph localization. (A) Synchronized RPE-1 cells were used to produce chromosome spreads, which were stained with antibodies specific for H2B S6ph (red) and CENP-A (green). Bar, 1 µm. The images were quantitatively analyzed with ImageJ for the distribution of H2B S6ph and CENP-A along the centromere cross section, as shown by the dotted line (top). The maximum of fluorescence intensity was set as 1, and shaded areas indicate standard deviations (n = 8 cells/368 centromere pairs). (B) The experiment was done as in Fig. 2 C, with the difference that chromosomes were stained for H2B S6ph and the cohesin component RAD21. Representative examples are shown; the boxed area is displayed in the bottom panel in 4× magnification. Bars: 5 µm (main); 1 µm (magnification). (C) Schematic representation of the chromosomal localization of H2B S6ph on mitotic chromatids.

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