Figure S4. Mini-nuclei in A. pullulans .... | Rockefeller University Press

Figure S4.

$Mini-nuclei in A. pullulans. (A) Inverted grayscale image of A. pullulans strains serially diluted and grown on YPD for 48 h at 24°C. (B) Representative maximum intensity projected confocal images of cells expressing 3xmCherry (cytosol, magenta) and NLS-GFP (nucleus, cyan) that have mini-nuclei (yellow arrows). (C) Histogram of the volume of the smallest nucleus in 625 multinucleate cells expressed as a percent of the volume of the largest nucleus for that cell. Red columns indicate cells where the smallest nuclear volume was <40% of the largest nuclear volume. (D) The fraction of cells with mini-nuclei, defined as nuclei with <40% the volume of the largest nucleus within the cell (n = 566, 333, 35, 99, 41, 22, 23, 34, 16, and 22 for cells with 1, 2, 3, 4, 5, 6, 7, 8, 9, and >9 nuclei). (E) Representative maximum intensity projected confocal image of S. cerevisiae (blue dashed outline) and A. pullulans (red dashed outline) SYTOX stained (cyan, DNA stain) cells in the same tube and imaged side by side. A mini-nucleus (yellow arrow) is visible in the A. pullulans cell. (F) A histogram showing the total SYTOX signal per S. cerevisiae and A. pullulans nucleus relative to the average SYTOX signal across all S. cerevisiae nuclei. (G) Representative maximum intensity projected confocal image of an A. pullulans cell expressing H3B-mCherry (magenta) with a mini-nucleus (yellow arrow). Scale bar in all images is 5 µm.$

Mini-nuclei in A. pullulans. (A) Inverted grayscale image of A. pullulans strains serially diluted and grown on YPD for 48 h at 24°C. (B) Representative maximum intensity projected confocal images of cells expressing 3xmCherry (cytosol, magenta) and NLS-GFP (nucleus, cyan) that have mini-nuclei (yellow arrows). (C) Histogram of the volume of the smallest nucleus in 625 multinucleate cells expressed as a percent of the volume of the largest nucleus for that cell. Red columns indicate cells where the smallest nuclear volume was <40% of the largest nuclear volume. (D) The fraction of cells with mini-nuclei, defined as nuclei with <40% the volume of the largest nucleus within the cell (n = 566, 333, 35, 99, 41, 22, 23, 34, 16, and 22 for cells with 1, 2, 3, 4, 5, 6, 7, 8, 9, and >9 nuclei). (E) Representative maximum intensity projected confocal image of S. cerevisiae (blue dashed outline) and A. pullulans (red dashed outline) SYTOX stained (cyan, DNA stain) cells in the same tube and imaged side by side. A mini-nucleus (yellow arrow) is visible in the A. pullulans cell. (F) A histogram showing the total SYTOX signal per S. cerevisiae and A. pullulans nucleus relative to the average SYTOX signal across all S. cerevisiae nuclei. (G) Representative maximum intensity projected confocal image of an A. pullulans cell expressing H3B-mCherry (magenta) with a mini-nucleus (yellow arrow). Scale bar in all images is 5 µm.

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