Journal: Nature Cell Biology
Article Title: Cytoplasmic flow is a cell size sensor that scales anaphase
doi: 10.1038/s41556-024-01605-6
Figure Lengend Snippet: a , Time lapse of a cell in anaphase (8-cell stage). H2B–mCherry, chromosomes; NLS-GFP and WGA-640, nucleus. Only one set of chromosomes is shown. t = 0 s, anaphase onset. WGA and NER are indicated. Scale bar, 20 µm. b , Kymograph of the cell in a . c , Time of WGA and NER (by NLS-GFP) as a function of cell length. t = 0 s, anaphase onset. n = 17, 14, 4, 7, 12, 16, 16 and 14 cells from 17, 12, 4, 5, 5, 5, 5 and 5 embryos at 4-, 8-, 16-, 32-, 64-, 128-, 256- and 512-cell stages, respectively. d , Position of WGA ( n = 17, 14, 4, 7, 12, 16, 16 and 12 cells from 17, 9, 3, 5, 5, 5, 5 and 5 embryos at 4-, 8-, 16-, 32-, 64-, 128-, 256- and 512-cell stages, respectively) and NER ( n = 17, 14, 3, 7, 12, 16, 15 and 13 cells from 17, 9, 3, 5, 5, 5, 5 and 5 embryos at 4-, 8-, 16-, 32-, 64-, 128-, 256- and 512-cell stages, respectively) as a function of cell length. e , Overlay of two timepoints for three developmental stages (4-, 32- and 128-cell stage), to visualize the scaling of NER. White, metaphase chromosomes; blue, chromosomes at NER. f , Cell size and position of NER in a 4-cell stage (yellow box; dashed line, cell boundary based on cytoplasmic NLS-GFP signal) and a 512-cell stage cell (white box). The NER distance in the large cell is longer than the cell size in the smaller cell. Scale bars, 10 µm. g , Semi-log plot of normalized pH3-s10 fluorescence levels for cells from 4- to 512-cell stages. The data are collapsed into a single exponential profile. n = 130 cells from 12 embryos, all developmental stages combined. h , Dephosphorylation rate of pH3-s10 (1/decay time) as a function of cell length. n = 9, 12, 11, 8, 14, 15, 15 and 14 cells from 9, 10, 10, 6, 9, 10, 9 and 7 embryos at 4-, 8-, 16-, 32-, 64-, 128-, 256- and 512-cell stages, respectively. i , The normalized pH3-s10 fluorescence intensity at WGA ( n = 2, 3, 2, 3, 5, 7, 10 and 12 cells from 2, 3, 2, 3, 2, 3, 4 and 4 embryos at 4-, 8-, 16-, 32-, 64-, 128-, 256- and 512-cell stages, respectively) and NER ( n = 2, 3, 2, 3, 5, 7, 10 and 10 cells from 2, 3, 2, 3, 2, 3, 4 and 4 embryos at 4-, 8-, 16-, 32-, 64-, 128-, 256- and 512-cell stages, respectively) as a function of cell length. j , Distance between the two sets of chromosomes as a function of time. Colours, developmental stages (4- to 512-stage). t = 0 s, anaphase onset. n = 35, 42, 23, 28, 39, 58, 45 and 40 cells from 35, 34, 22, 22, 21, 23, 17 and 13 embryos at 4-, 8-, 16-, 32-, 64-, 128-, 256- and 512-cell stages, respectively. k , Chromosome separation velocity as a function of cell length. v A (0–100 s) and \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${v}_{{\mathrm{B}}}$$\end{document} v B (100–200 s after anaphase onset). For \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${v}_{{\mathrm{A}}}$$\end{document} v A , n = 26, 30, 17, 23, 35, 50, 43 and 40 cells from 26, 24, 16, 16, 17, 18, 17 and 13 embryos at 4-, 8-, 16-, 32-, 64-, 128-, 256- and 512-cell stages, respectively. For \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${v}_{{\mathrm{B}}}$$\end{document} v B , n = 34, 41, 20, 24, 39, 58, 45 and 40 cells from 34, 34, 19, 19, 21, 23, 17 and 13 embryos at 4-, 8-, 16-, 32-, 64-, 128-, 256- and 512-cell stages, respectively. l , m , Kymographs of a big and small cell (4- and 128-cell stages, respectively) showing chromosomes ( l ) and WGA and NER ( m ). The dashed white line in l represents chromosome velocity. \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${v}_{A}$$\end{document} v A is the same for both cells, whereas \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${v}_{B}$$\end{document} v B changes. In c , d , h , i and k , the bins represent the cell stage. In a – g , i and k – m , the error bars represent s.d. In h and j , the error bars represent s.e.m.
Article Snippet: The primary antibody was rabbit anti-pH3-s10 D2C8 (1:200; 3377; Cell Signalling (lot 7)), and the secondary antibody was anti-rabbit Alexa-488 (1:200).
Techniques: Fluorescence, De-Phosphorylation Assay
Bethyl is a verified supplier 
![a , Time lapse of a cell in anaphase (8-cell stage). H2B–mCherry, chromosomes; NLS-GFP and WGA-640, nucleus. Only one set of chromosomes is shown. t = 0 s, anaphase onset. WGA and NER are indicated. Scale bar, 20 µm. b , Kymograph of the cell in a . c , Time of WGA and NER (by NLS-GFP) as a function of cell length. t = 0 s, anaphase onset. n = 17, 14, 4, 7, 12, 16, 16 and 14 cells from 17, 12, 4, 5, 5, 5, 5 and 5 embryos at 4-, 8-, 16-, 32-, 64-, 128-, 256- and 512-cell stages, respectively. d , Position of WGA ( n = 17, 14, 4, 7, 12, 16, 16 and 12 cells from 17, 9, 3, 5, 5, 5, 5 and 5 embryos at 4-, 8-, 16-, 32-, 64-, 128-, 256- and 512-cell stages, respectively) and NER ( n = 17, 14, 3, 7, 12, 16, 15 and 13 cells from 17, 9, 3, 5, 5, 5, 5 and 5 embryos at 4-, 8-, 16-, 32-, 64-, 128-, 256- and 512-cell stages, respectively) as a function of cell length. e , Overlay of two timepoints for three developmental stages (4-, 32- and 128-cell stage), to visualize the scaling of NER. White, metaphase chromosomes; blue, chromosomes at NER. f , Cell size and position of NER in a 4-cell stage (yellow box; dashed line, cell boundary based on cytoplasmic NLS-GFP signal) and a 512-cell stage cell (white box). The NER distance in the large cell is longer than the cell size in the smaller cell. Scale bars, 10 µm. g , Semi-log plot of normalized <t>pH3-s10</t> fluorescence levels for cells from 4- to 512-cell stages. The data are collapsed into a single exponential profile. n = 130 cells from 12 embryos, all developmental stages combined. h , Dephosphorylation rate of pH3-s10 (1/decay time) as a function of cell length. n = 9, 12, 11, 8, 14, 15, 15 and 14 cells from 9, 10, 10, 6, 9, 10, 9 and 7 embryos at 4-, 8-, 16-, 32-, 64-, 128-, 256- and 512-cell stages, respectively. i , The normalized pH3-s10 fluorescence intensity at WGA ( n = 2, 3, 2, 3, 5, 7, 10 and 12 cells from 2, 3, 2, 3, 2, 3, 4 and 4 embryos at 4-, 8-, 16-, 32-, 64-, 128-, 256- and 512-cell stages, respectively) and NER ( n = 2, 3, 2, 3, 5, 7, 10 and 10 cells from 2, 3, 2, 3, 2, 3, 4 and 4 embryos at 4-, 8-, 16-, 32-, 64-, 128-, 256- and 512-cell stages, respectively) as a function of cell length. j , Distance between the two sets of chromosomes as a function of time. Colours, developmental stages (4- to 512-stage). t = 0 s, anaphase onset. n = 35, 42, 23, 28, 39, 58, 45 and 40 cells from 35, 34, 22, 22, 21, 23, 17 and 13 embryos at 4-, 8-, 16-, 32-, 64-, 128-, 256- and 512-cell stages, respectively. k , Chromosome separation velocity as a function of cell length. v A (0–100 s) and \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${v}_{{\mathrm{B}}}$$\end{document} v B (100–200 s after anaphase onset). For \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${v}_{{\mathrm{A}}}$$\end{document} v A , n = 26, 30, 17, 23, 35, 50, 43 and 40 cells from 26, 24, 16, 16, 17, 18, 17 and 13 embryos at 4-, 8-, 16-, 32-, 64-, 128-, 256- and 512-cell stages, respectively. For \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${v}_{{\mathrm{B}}}$$\end{document} v B , n = 34, 41, 20, 24, 39, 58, 45 and 40 cells from 34, 34, 19, 19, 21, 23, 17 and 13 embryos at 4-, 8-, 16-, 32-, 64-, 128-, 256- and 512-cell stages, respectively. l , m , Kymographs of a big and small cell (4- and 128-cell stages, respectively) showing chromosomes ( l ) and WGA and NER ( m ). The dashed white line in l represents chromosome velocity. \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${v}_{A}$$\end{document} v A is the same for both cells, whereas \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${v}_{B}$$\end{document} v B changes. In c , d , h , i and k , the bins represent the cell stage. In a – g , i and k – m , the error bars represent s.d. In h and j , the error bars represent s.e.m.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_1524/pmc11821524/pmc11821524__41556_2024_1605_Fig1_HTML.jpg)
