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. 2018 Mar 26;7:e34122. doi: 10.7554/eLife.34122

Figure 5. D1 bundles satellite DNA from heterologous chromosomes to form chromocenter.

Figure 5.

(A, B) C2C12 cells expressing GFP only (blue) (A) or GFP-D1 (blue) (B) stained for DAPI (red). Dotted lines indicate nucleus. (C) Quantification of chromocenter number relative to expression level of GFP (n = 29) or GFP-D1 (n = 47). P value and R2 value are indicated from linear regression analysis. (D) FISH against LacO (red) and {AATAT}n (green) on mitotic neuroblast chromosomes from the LacO strain stained for DAPI (blue), indicating the sites of LacO insertion (arrows). (E, F) FISH against LacO (red) and {AATAT}n (green) in spermatogonia expressing GFP-D1 (blue) (E) or GFP-LacI-D1 (blue) (F). Arrows indicate location of LacO sequence. (G) AATAT-LacO distance (nm) in GFP-D1 (n = 97) and GFP-LacI-D1 (n = 69) expressing spermatogonia. P value from student’s t-test is shown. Error bars: SD. All scale bars: 5 μm.

Figure 5—source data 1. Quantification of the relative expression of GFP/GFP-D1 and number of chromocenters in mouse cells.
Images of cells expressing GFP (n=27) or GFP-D1 (n=47) were acquired from a single slide/transfection with all image acquisition parameters held constant and care taken to avoid pixel oversaturation. Total GFP fluorescence per cell was quantified using ImageJ software. Relative expression was calculated by dividing each individual fluorescence value over the maximum fluorescence value obtained over the course of the experiment. DAPI staining was used to calculate number of chromocenters per cell.
elife-34122-fig5-data1.xlsx (9.7KB, xlsx)
DOI: 10.7554/eLife.34122.014
Figure 5—source data 2. Quantification of LacO-AATAT distance (nm) in cells expressing GFP-D1 and GFP-LacI-D1.
LacO-AATAT distance (nm) was measured in spermatogonial cells expressing GFP-D1 (n=97) and GFP-LacI-D1 (n=69) using Leica LAS X software.
elife-34122-fig5-data2.xlsx (10.3KB, xlsx)
DOI: 10.7554/eLife.34122.015