Skip to main content
. Author manuscript; available in PMC: 2019 Nov 5.
Published in final edited form as: Dev Cell. 2018 Oct 4;47(3):348–362.e7. doi: 10.1016/j.devcel.2018.09.003

Figure 1. Labeling of proteins transiently associated with CENP-A and H3.1 nucleosomes.

Figure 1.

(A) Schematic representation of the experimental approach. (B) (C) Representative images of cells stably expressing indicated proteins fused to the BirA* ligase and HA tag, and incubated with medium supplemented with or without biotin for 5 hours. DNA is visualized by DAPI staining, immunofluorescence for CENP-T is shown in red, biotinylated proteins (B) or BirA*-HA fusion proteins (C) are shown in green. Scale bar is 5μm. (D) Streptavidin purification of biotinylated proteins from indicated cells lines analyzed by immunoblot. Cell media was supplemented with or without biotin for 24 hours. (E) Graph showing SILAC comparison between CENP-A-BirA*-HA biotinylation profile (heavy) versus biotinylation profile of H3.1-BirA-HA* (light) in cells undergoing S phase. Corresponding H/L scores for selected proteins are displayed. The graph represents an average of two independently performed experiments +/− SEM.