Disrupting the H3pT3–Survivin interaction reveals an H2ApT120-dependent pool of Aurora B at the KT-proximal centromere. (A) DMSO or BAY 1816032 was added to the indicated Survivin stable cell lines 9 h after release from STB, and then MG132 was added 10 h after release. Chromosome spreads were prepared from mitotic cells collected 15 h after release and then immunostained. (B) DMSO or the indicated inhibitors was added to HeLa cells 7 h after release from STB, and MG132 was added 10 h after release. Chromosome spreads were prepared and immunostained as described in A. (C) Nocodazole and DMSO or the indicated inhibitors were added to HeLa cells 9 h after release from STB. Chromosome spreads were prepared from mitotic cells collected 12 h after release and then immunostained. Line plots and scatter plots show the fluorescence intensity across a sister kinetochore pair for Aurora B and the inner kinetochore marker CENP-C on 1 chromosome and 10 chromosomes, respectively. (D) Nocodazole and DMSO or the indicated inhibitors were added to HeLa cells 9 h after release from STB. Chromosome spreads were prepared and immunostained as described in C. Line plots and scatter plots show the fluorescence intensity across a sister kinetochore pair for Aurora B and H2ApT120 on 1 chromosome and 10 chromosomes, respectively. Scale bars, 10 µm.