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. 2017 Jan 31;27(4):483–504. doi: 10.1038/cr.2017.18

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Copyright © 2017 Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences

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TTALE-mediated imaging of telomeres and centromeres during human stem cell aging. (A) Schematic diagram of three established human stem cell aging models. (B) SIM images showing EGFP-TTALE^telo-labeled telomeres in WS-MSCs vs WT-MSCs at passage 6 (top), HGPS-MSCs vs HGPS-GC-MSCs at passage 8 (middle), and LP-WT-MSCs at passage 12 vs EP-WT-MSCs at passage 6 (bottom). Magnified images of the boxed regions are in the lower-left corner. Dashed lines indicate the nuclear boundary. Scale bars, 5 μm (wide-field images) or 10 μm (magnified images). (C) Decrease of EGFP-TTALE^telo fluorescence intensity with hMSC aging. The scatter plot displays the fluorescence intensity of EGFP-TTALE^telo-labeled telomere puncta normalized by the average fluorescence intensity of the nucleus in the indicated cells. n = 50 nuclei per group; ^***P < 0.001. (D) SIM images of centromeres labeled by EGFP-TTALE^centro in WS-MSCs and WT-MSCs. Bottom panels show magnified images of boxed regions from the top panels. Dashed lines indicate the nuclear boundary. Scale bars, 5 μm. (E) Intensity profiles of TTALE^centro signals across the dotted lines (12 μm in length) in the bottom panels of D. (F) Histogram showing the percentage of fluorescence-labeled area relative to nuclear area in the indicated cells. n = 50 nuclei per group; ^***P < 0.001.