Figure 3.
KAT7 and Other MYST Subunits Localize to the Centromere
(A) KAT7 and CENP-A co-localization. Fluorescence images were obtained with DAPI (DNA), EYFP (green), anti-CENP-A antibody (red), and anti-cyclin B antibody. Cell cycles were distinguished by images of DAPI and cyclin B staining, and these distributions are shown in the upper right panel. The observed frequency of KAT7 centromere localization positive cells (i.e., co-localization of KAT7 and CENP-A) is plotted in the bottom right panel. EYFP-BRPF2, -ING4, and -ING5 were also analyzed. See also Figure S3B.
(B) EYFP-KAT7 localized to centromeres at an earlier stage of G1 phase, using the presence of an intercellular bridge of tubulin as a marker for early G1 cells. Fluorescence images were obtained with DAPI (DNA), EYFP (green), anti-CENP-A antibody (red), and anti-tubulin antibody (blue).
(C) KAT7 co-localization with new CENP-A and Halo-HJURP in G1 cells. (Left) Scheme for a quench chase pulse labeling of SNAP-CENP-A in G1 phase or G1 cell observation is shown on the left. Fluorescence images were obtained with DAPI (DNA), EYFP (green), TMR ligand (red; TMR-Star or Halo-TMR), and anti-CENP-A antibody. Data in this figure are presented as mean ± SD. (n = 3, >50 cells counted in each observation).
Scale bars, 5 μm.