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. 2015 May 15;29(10):1058–1073. doi: 10.1101/gad.259432.115

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© 2015 Fang et al.; Published by Cold Spring Harbor Laboratory Press

This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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Figure 4. — Specific loading of CENP-N in middle/late S phase (see also Supplemental Fig. S6). (A) Representative images of HeLa cells expressing SNAP-CENP-N in different cell cycle phases. SNAP-CENP-N was labeled with TMR-Star (red), and cell cycle phases were determined by combined staining with CENP-F antibody (gray), PCNA antibody (green), and DAPI (blue). Bar, 10 µm. (B) Statistical analysis of the relative fluorescence intensity of SNAP-CENP-N during the cell cycle as shown in A. (C) Outline of cell synchronization and the labeling regimen for CENP-N loading. (D) HeLa cells expressing SNAP-CENP-N (red) were synchronized and labeled as depicted in C, and cell cycle phases were determined by combined staining with CENP-F antibody (gray), PCNA antibody (green), and DAPI (blue). Bar, 10 µm. (E) Statistical analysis of relative fluorescence intensity of SNAP-CENP-N newly synthesized and deposited onto centromeres as shown in D.