Figure 1.
The centromere is rapidly disassembled upon quiescence entry. (A) Graph showing CENP-T centromere intensity level over time of quiescence entry. Each point indicates the average centromere intensity level for all centromeres in a single cell, adjusted for background. Intensity values were normalized to day 0. Red line represents the median; blue points represent average of each replicate. Points were aggregated from three replicates. n = 224, 392, 386, 419, 310, and 410 cells for 0, 1, 2, 3, 5, and 7-day time points, respectively. P values can be found in Table S2. (B) Representative immunofluorescence images of a cycling cell and cell in quiescence for 7 days. Cells were stained with CENP-T and anti-centromere (ACA) antibodies. Scale bar = 5 µm. (C) Graph showing CENP-L centromere intensity level over time of quiescence entry. Each point indicates the average centromere intensity level for all centromeres in a single cell, adjusted for background. Intensity values were normalized to day 0. Red line represents the median, and blue points represent average of each replicate. Points were aggregated from three replicates. n = 224, 300, 314, 288, 281, and 290 cells for 0, 1, 2, 3, 5, and 7-day time points, respectively. P values can be found in Table S2. (D) Representative immunofluorescence images of a cycling cell and cell in quiescence for 7 days. Cells were stained with CENP-L and anti-centromere (ACA) antibodies. Scale bar = 5 µm. (E) Graph showing CENP-K centromere intensity level over time of quiescence entry. Each point indicates the average centromere intensity level for all centromeres in a single cell, adjusted for background. Intensity values were normalized to day 0. Red line represents the median, and blue points represent average of each replicate. Points were aggregated from three replicates. n = 162, 442, 482, 525, 470, and 355 cells for 0, 1, 2, 3, 5, and 7-day time points, respectively. P values can be found in Table S2. (F) Representative immunofluorescence images of a cycling cell and cell in quiescence for 7 days. Cells were stained with CENP-K and anti-centromere (ACA) antibodies. Scale bar = 5 µm. (G) Graph showing CENP-O/P centromere intensity level over time of quiescence entry. Each point indicates the average centromere intensity level for all centromeres in a single cell, adjusted for background. Intensity values were normalized to day 0. Red line represents the median, and blue points represent average of each replicate. Points were aggregated from three replicates. n = 181, 522, 514, 465, 467, and 454 cells for 0, 1, 2, 3, 5, and 7-day time points respectively. P values can be found in Table S2. (H) Representative immunofluorescence images of a cycling cell and cell in quiescence for 7 days. Cells were stained with CENP-O/P and anti-centromere (ACA) antibodies. Scale bar = 5 µm. (I) Graph showing CENP-C centromere intensity level over time of quiescence entry. Each point indicates the average centromere intensity level for all centromeres in a single cell, adjusted for background. Intensity values were normalized to day 0. Red line represents the median, and blue points represent average of each replicate. Points were aggregated from three or more replicates. n = 201, 174, 169, 166, 196, and 194 cells for 0, 1, 2, 3, 5, and 7-day time points, respectively. P values can be found in Table S2. (J) Representative immunofluorescence images of a cycling cell and cell in quiescence for 7 days. Cells were stained with CENP-C and anti-centromere (ACA) antibodies. Scale bar = 5 µm. (K) Western blots of cells in quiescence for the indicated amount of days. Blots were incubated with CENP-T and CENP-H antibodies. β-Actin is used as a loading control. (L) Western blot of cells in quiescence for the indicated amount of days. Blot was incubated with CENP-C antibody. β-Actin is used as a loading control. Source data are available for this figure: SourceData F1. Refer to the image caption for details. Panel A: A scatter plot showing C E N P-T centromere intensity over days in quiescence. The horizontal axis represents days in quiescence, and the vertical axis represents centromere intensity. The red line indicates the median intensity, and blue points represent the average of each replicate. Panel B: Immunofluorescence images of cycling and quiescent cells stained with C E N P-T and anti-centromere antibodies. Panel C: A scatter plot showing C E N P-L centromere intensity over days in quiescence. The horizontal axis represents days in quiescence, and the vertical axis represents centromere intensity. The red line indicates the median intensity, and blue points represent the average of each replicate. Panel D: Immunofluorescence images of cycling and quiescent cells stained with C E N P-L and anti-centromere antibodies. Panel E: A scatter plot showing C E N P-K centromere intensity over days in quiescence. The horizontal axis represents days in quiescence, and the verytical axis represents centromere intensity. The red line indicates the median intensity, and blue points represent the average of each replicate. Panel F: Immunofluorescence images of cycling and quiescent cells stained with C E N P-K and anti-centromere antibodies. Panel G: A scatter plot showing C E N P-O over P centromere intensity over days in quiescence. The horizontal axis represents days in quiescence, and the vertical axis represents centromere intensity. The red line indicates the median intensity, and blue points represent the average of each replicate. Panel H: Immunofluorescence images of cycling and quiescent cells stained with C E N P-O over P and anti-centromere antibodies. Panel I: A scatter plot showing C E N P-C centromere intensity over days in quiescence. The horizontal axis represents days in quiescence, and the vertical axis represents centromere intensity. The red line indicates the median intensity, and blue points represent the average of each replicate. Panel J: Immunofluorescence images of cycling and quiescent cells stained with C E N P-C and anti-centromere antibodies. Panel K: Western blot images showing C E N P-T and C E N P-H protein levels in quiescent cells over days. Beta-actin is used as a loading control. Panel L: Western blot image showing C E N P-C protein levels in quiescent cells over days. Beta-actin is used as a loading control. All data is approximate.

The centromere is rapidly disassembled upon quiescence entry. (A) Graph showing CENP-T centromere intensity level over time of quiescence entry. Each point indicates the average centromere intensity level for all centromeres in a single cell, adjusted for background. Intensity values were normalized to day 0. Red line represents the median; blue points represent average of each replicate. Points were aggregated from three replicates. n = 224, 392, 386, 419, 310, and 410 cells for 0, 1, 2, 3, 5, and 7-day time points, respectively. P values can be found in Table S2. (B) Representative immunofluorescence images of a cycling cell and cell in quiescence for 7 days. Cells were stained with CENP-T and anti-centromere (ACA) antibodies. Scale bar = 5 µm. (C) Graph showing CENP-L centromere intensity level over time of quiescence entry. Each point indicates the average centromere intensity level for all centromeres in a single cell, adjusted for background. Intensity values were normalized to day 0. Red line represents the median, and blue points represent average of each replicate. Points were aggregated from three replicates. n = 224, 300, 314, 288, 281, and 290 cells for 0, 1, 2, 3, 5, and 7-day time points, respectively. P values can be found in Table S2. (D) Representative immunofluorescence images of a cycling cell and cell in quiescence for 7 days. Cells were stained with CENP-L and anti-centromere (ACA) antibodies. Scale bar = 5 µm. (E) Graph showing CENP-K centromere intensity level over time of quiescence entry. Each point indicates the average centromere intensity level for all centromeres in a single cell, adjusted for background. Intensity values were normalized to day 0. Red line represents the median, and blue points represent average of each replicate. Points were aggregated from three replicates. n = 162, 442, 482, 525, 470, and 355 cells for 0, 1, 2, 3, 5, and 7-day time points, respectively. P values can be found in Table S2. (F) Representative immunofluorescence images of a cycling cell and cell in quiescence for 7 days. Cells were stained with CENP-K and anti-centromere (ACA) antibodies. Scale bar = 5 µm. (G) Graph showing CENP-O/P centromere intensity level over time of quiescence entry. Each point indicates the average centromere intensity level for all centromeres in a single cell, adjusted for background. Intensity values were normalized to day 0. Red line represents the median, and blue points represent average of each replicate. Points were aggregated from three replicates. n = 181, 522, 514, 465, 467, and 454 cells for 0, 1, 2, 3, 5, and 7-day time points respectively. P values can be found in Table S2. (H) Representative immunofluorescence images of a cycling cell and cell in quiescence for 7 days. Cells were stained with CENP-O/P and anti-centromere (ACA) antibodies. Scale bar = 5 µm. (I) Graph showing CENP-C centromere intensity level over time of quiescence entry. Each point indicates the average centromere intensity level for all centromeres in a single cell, adjusted for background. Intensity values were normalized to day 0. Red line represents the median, and blue points represent average of each replicate. Points were aggregated from three or more replicates. n = 201, 174, 169, 166, 196, and 194 cells for 0, 1, 2, 3, 5, and 7-day time points, respectively. P values can be found in Table S2. (J) Representative immunofluorescence images of a cycling cell and cell in quiescence for 7 days. Cells were stained with CENP-C and anti-centromere (ACA) antibodies. Scale bar = 5 µm. (K) Western blots of cells in quiescence for the indicated amount of days. Blots were incubated with CENP-T and CENP-H antibodies. β-Actin is used as a loading control. (L) Western blot of cells in quiescence for the indicated amount of days. Blot was incubated with CENP-C antibody. β-Actin is used as a loading control. Source data are available for this figure: SourceData F1.

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