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. 2021 Oct 2;10(10):2638. doi: 10.3390/cells10102638

Figure 6.

Figure 6

MIBS assembly is regulated by CDK1 activity. (A) Maximum intensity projections of confocal image stacks from representative HeLa-RFP-H2B cells synchronized in mitosis by a double thymidine block, showing staining of p62, β-tubulin, and DNA (Hoechst); cells were treated or not with low doses of the CDK1 inhibitor RO3306 for 3 h before cell fixation. Enlarged views of boxed regions are shown to emphasize MIBS assembly until metaphase and their disassembly at anaphase, and inhibition of MIBS assembly by the CDK1 inhibitor. A heatmap pseudo-color intensity scale was applied to the p62 staining channel to underscore MIBS assembly dynamics; Bars: 10 µm or 5 µm. (B) Quantification of cells from (A), indicating MIBS number per cell and their mean size during mitotic progression, means ± SE from 11 to 59 cells from 2 representative experiments. Statistical significance was analyzed with the Kruskal–Wallis test: ****, p < 0.0001; ***, p < 0.001; **: p < 0.005; ns, not significant. (C) Schematic of the protocol used. Quantification of cells from (A), showing percentages of cells treated with RO3306 or the vehicle (DMSO) at distinct mitotic stages, means ± SE from 411 to 438 cells from 2 representative experiments.