Figure 1. Localization of cohesin and... | Rockefeller University Press

$Figure 1. Localization of cohesin and condensin in the pericentric chromatin. (A) Smc3-GFP (cohesin) and Smc4-GFP (condensin) are enriched in the metaphase spindle between spindle pole bodies (Spc29-RFP). A line scan (dotted yellow line) of Smc3-GFP perpendicular to the spindle axis shows that cohesin has a bilobed enrichment. Line scans of Smc4-GFP taken along the spindle axis and between spindle pole bodies revealed three distinct classes of pericentric condensin enrichment: one focus, two foci, and uniform distribution left to right (n = 80, with percentages listed). Condensin ribosomal DNA localization is labeled with a white arrow. (B) Transverse (end on) images of Smc3-GFP display an area of low fluorescence and are displaced from the spindle pole. Smc4-GFP displays a diffraction-limited spot in line with the spindle pole body. Vertical line scans are shown as graphs below their respective images. (A and B) The schematics depict the image viewing angle of the mitotic spindle. In A, the spindle is viewed from the side/sagittal with both spindle poles in focus. In B, the spindle is viewed end on/transverse, and the spindle poles are aligned on top of each other so that only one pole is visible. (C) Ensemble-averaged images of Smc3-GFP, Smc4-GFP, and Nuf2-GFP (kinetochore protein) were generated by aligning and scaling multiple single-plane images of each protein. Line scans of single-plane images were averaged and graphed to quantify the distribution. The error bars represent SEM (Smc3, n = 34; Smc4, n = 51; Nuf2, n = 30). (D) Images of Smc3 and Smc4 relative to the kinetochore (Ndc80) and spindle poles (Spc29; white arrows). Line scans are shown as graphs below their respective image. All line scan graphs display the relative fluorescence intensity plotted versus the distance in pixels (65 nm/pixel). Bars, 1 µm.$

Figure 1.

Localization of cohesin and condensin in the pericentric chromatin. (A) Smc3-GFP (cohesin) and Smc4-GFP (condensin) are enriched in the metaphase spindle between spindle pole bodies (Spc29-RFP). A line scan (dotted yellow line) of Smc3-GFP perpendicular to the spindle axis shows that cohesin has a bilobed enrichment. Line scans of Smc4-GFP taken along the spindle axis and between spindle pole bodies revealed three distinct classes of pericentric condensin enrichment: one focus, two foci, and uniform distribution left to right (n = 80, with percentages listed). Condensin ribosomal DNA localization is labeled with a white arrow. (B) Transverse (end on) images of Smc3-GFP display an area of low fluorescence and are displaced from the spindle pole. Smc4-GFP displays a diffraction-limited spot in line with the spindle pole body. Vertical line scans are shown as graphs below their respective images. (A and B) The schematics depict the image viewing angle of the mitotic spindle. In A, the spindle is viewed from the side/sagittal with both spindle poles in focus. In B, the spindle is viewed end on/transverse, and the spindle poles are aligned on top of each other so that only one pole is visible. (C) Ensemble-averaged images of Smc3-GFP, Smc4-GFP, and Nuf2-GFP (kinetochore protein) were generated by aligning and scaling multiple single-plane images of each protein. Line scans of single-plane images were averaged and graphed to quantify the distribution. The error bars represent SEM (Smc3, n = 34; Smc4, n = 51; Nuf2, n = 30). (D) Images of Smc3 and Smc4 relative to the kinetochore (Ndc80) and spindle poles (Spc29; white arrows). Line scans are shown as graphs below their respective image. All line scan graphs display the relative fluorescence intensity plotted versus the distance in pixels (65 nm/pixel). Bars, 1 µm.

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