Fig. 7: RSF1 localization to mitotic kinetochores requires the phosphorylation of histone H3 on Ser28.

a HeLa cells were treated with nocodazole or paclitaxel for 4 h and prometaphase cells were stained with anti-RSF1 (green) and anti-Aurora B (red) antibodies. Scale bars, 5 μm. b Nocodazole- or paclitaxel-treated HeLa cells were immunoprecipitated using anti-RSF1 antibodies. The inputs and immunoprecipitants were analyzed by western blotting. c HeLa cells were treated with paclitaxel treated for 4 h and prometaphase cells were pre-treated with Aurora B inhibitor ZM447439 for 30 min in the presence of MG132. Mitotic HeLa cells were stained with anti-RSF1 (green) and anti-BubR1 (red) antibodies. Scale bars, 5 μm. d HeLa cells were transfected with indicated GFP-Histone H3 WT or S28A/D constructs and treated with paclitaxel for 16 h. Mitotic cell lysates were followed by immunoprecipitation using anti-RSF1 antibody. e HeLa cells were introduced with GFP-Histone H3 WT or S28A/D mutants and treated with paclitaxel for 4 h. Images were obtained from representative mitotic cells: RSF1 (green), Aurora B (red), and GFP (gray). Scale bars, 5 μm. f A proposed model for spatiotemporal regulation of the on/off switch on Aurora B kinase by the RSF1-PLK1 axis. New findings are colored in orange lines. Data of b, d are representative of three independent experiments. Source data are provided as a Source Data file.