Extended Data Fig. 6: Chromosome clustering is not mediated by association with BAF or removal of Ki-67.
From: Chromosome clustering by Ki-67 excludes cytoplasm during nuclear assembly
a–c, Effect of BAF depletion on chromosome clustering. a, Live mitotic HeLa cells stably expressing H2B–mCherry and GEMs were imaged 72 h after siRNA transfection, in the presence of nocodazole; flavopiridol was added at t = 0 min to induce mitotic exit. White dashed lines indicate chromosomal areas, single z-slice shown. b, Quantification of chromosome convex hull area, normalized to pre-flavopiridol addition. c, GEM particle count within chromosomal area normalized to pre-flavopiridol addition. n = 23 cells (siBAF), n = 23 cells (siControl). d, Immunoblot analysis of BAF and actin 72 h after siRNA transfection, showing one of two biological replicates. For gel source data, see Supplementary Fig. 1. e, Localization of BAF–eGFP in live mitotic HeLa cell imaged in the presence of nocodazole; flavopiridol was added at t = 0 s to induce mitotic exit. f, Quantification of chromosome convex hull area and BAF–eGFP accumulation at the surface of the chromatin region as in e, normalized to pre-flavopiridol. n = 21 cells. g, Localization of Ki-67, eGFP tagged on endogenous loci in live HeLa cell progressing from metaphase to anaphase (anaphase onset = 0 min), DNA was stained with SiR-Hoechst, z-projection. h, Quantification of chromosome convex hull area and eGFP–Ki-67 mean fluorescence intensity on chromosomes, normalized to pre-anaphase, for 41 cells as in g. i, Localization of Ki-67 during spindle-less mitotic exit. Time-lapse microscopy of mitotic HeLa cell as in g, in the presence of nocodazole; flavopiridol was added (t = 0 min) to induce mitotic exit. Z-projection. j, Quantification of chromosome convex hull area and eGFP–Ki-67 mean fluorescence intensity on chromosomes of 27 cells as in i. Lines and shaded areas indicate mean ± s.d., scale bars, 10 μm.
