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. 2022 Aug 3;609(7925):183–190. doi: 10.1038/s41586-022-05027-y

Fig. 1. Acetylation-regulated chromatin compaction prevents microtubule perforation in mitosis.

Fig. 1

a, The contribution of condensin and histone deacetylases to mitotic chromosome compaction and congression to the spindle centre. HeLa cells with homozygously mAID-tagged SMC4 were treated with 5-PhIAA to deplete condensin (ΔCondensin) or with TSA to suppress mitotic histone deacetylation as indicated. Live-cell images with microtubules stained by SiR–tubulin; DNA was stained with Hoechst 33342. Projection of 5 z-sections. b, Quantification of chromosome congression by the fraction of chromatin localizing to the central spindle region. n = 51 (control), n = 65 (ΔCondensin), n = 34 (ΔCondensin + TSA), n = 61 (TSA) cells. The bars indicate the mean. Significance was tested using two-tailed Mann–Whitney U-tests (P < 10−15 (ΔCondensin + TSA); P < 10−15 (TSA); precision limit of floating-point arithmetic). c, Quantification of chromatin density in cells treated as described in a. n = 31 (control), n = 89 (ΔCondensin), n = 99 (ΔCondensin + TSA) and n = 74 (TSA) cells. The bars indicate the mean. Significance was tested using two-tailed Mann–Whitney U-tests (P < 10−15 (ΔCondensin + TSA); P < 10−15 (TSA); precision limit of floating-point arithmetic). AU, arbitrary units. d, Electron tomography analysis of wild-type prometaphase HeLa cells in the absence or presence of TSA. Magenta, chromatin surfaces; green, microtubules in cytoplasm; cyan, microtubules in chromatin. The red circles show the perforation sites. e,f, Quantification of microtubule density in chromatin (e) and cytoplasmic (f) regions as shown in d. n = 10 tomograms from 7 cells for each condition. The bars indicate the mean. Significance was tested using two-tailed Mann–Whitney U-tests (P = 1.083 × 10−5 (e); P = 0.247 (f)). Biological replicates: n = 2 (af). Scale bars, 5 µm (a), 2 µm (d, 250 nm section); 200 nm (tomogram slices and 3D model).

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