Figure 2.
Disrupting the H3pT3–Survivin interaction increases H2ApT120 and Sgo1 at metaphase centromeres.(A–E) HeLa cells stably expressing the indicated Survivin proteins were released from STB for 12 h and then immunostained. Example images are shown (A). For each experiment, the immunofluorescence intensity ratios of H2ApT120/ACA (B) and Sgo1/ACA (D) were determined from 200 chromosomal regions containing >600 centromeres in 20 cells. The relative intensities of H2ApT120 (C) and Sgo1 (E), representing the normalized ratios of H2ApT120/ACA and Sgo1/ACA, were determined in 60 cells from three independent experiments (20 cells per experiment). (F–J) DMSO or 5-ITu was added to HeLa cells 9 h after release from STB. Cells were then fixed 12 h after release for immunostaining. Example images are shown (F). For each experiment, the immunofluorescence intensity ratios of H2ApT120/ACA (G) and Sgo1/ACA (I) were determined from 200 chromosomal regions containing >600 centromeres in 20 cells. The relative intensities of H2ApT120 (H) and Sgo1 (J), representing the normalized ratios of H2ApT120/ACA and Sgo1/ACA, were determined in 60 cells from three independent experiments (20 cells per experiment). Means and SDs are shown (B–E and G–J; unpaired t test). N.S., not significant. Scale bars, 10 µm.

Disrupting the H3pT3–Survivin interaction increases H2ApT120 and Sgo1 at metaphase centromeres. (A–E) HeLa cells stably expressing the indicated Survivin proteins were released from STB for 12 h and then immunostained. Example images are shown (A). For each experiment, the immunofluorescence intensity ratios of H2ApT120/ACA (B) and Sgo1/ACA (D) were determined from 200 chromosomal regions containing >600 centromeres in 20 cells. The relative intensities of H2ApT120 (C) and Sgo1 (E), representing the normalized ratios of H2ApT120/ACA and Sgo1/ACA, were determined in 60 cells from three independent experiments (20 cells per experiment). (F–J) DMSO or 5-ITu was added to HeLa cells 9 h after release from STB. Cells were then fixed 12 h after release for immunostaining. Example images are shown (F). For each experiment, the immunofluorescence intensity ratios of H2ApT120/ACA (G) and Sgo1/ACA (I) were determined from 200 chromosomal regions containing >600 centromeres in 20 cells. The relative intensities of H2ApT120 (H) and Sgo1 (J), representing the normalized ratios of H2ApT120/ACA and Sgo1/ACA, were determined in 60 cells from three independent experiments (20 cells per experiment). Means and SDs are shown (B–E and G–J; unpaired t test). N.S., not significant. Scale bars, 10 µm.

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