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. 2022 Jun 30;29(7):639–652. doi: 10.1038/s41594-022-00790-y

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Extended Data Fig. 2 — (a) Cell counts of an additional RPE FAP-TRF1 clone (#18) obtained 4 days after recovery from indicated treatments. (b) Cell counts of RPE FAP-TRF1 (red) and BJ FAP-TRF1 (blue) cells 4 days after recovery from dye + light treatments relative to untreated. Error bars represent the means ± s.d. from three independent experiments. (c-d) Cell counts of parental BJ-hTERT (c) or RPE-hTERT (d) cells obtained 4 days after recovery from indicated treatments. (e) Cell counts of FAP-TRF1 expressing HeLa and U2OS clones obtained 4 days after recovery from 5 min dye + light treatment relative to untreated. Black circles indicate the number of independent experiments. (f) Representative image of FAP-mCER-TRF1 protein colocalization with telomeres in bulk population primary BJ cells visualized by mCER IF (pink) with telo-FISH (green). (g-h) Cell counts of BJ (g) or RPE (h) FAP-TRF1 cells 4 days after recovery from one hour treatments with 2.5 or 10 mM KBrO₃, and for BJ FAP-TRF1 with 50 μM etoposide (ETP). (i) Cell counts of BJ or RPE FAP-TRF1 cells obtained 24 hours after recovery from 5 min dye + light treatment relative to untreated. For panels a, c-d and g-i, error bars represent the mean ± s.d. from the number of independent experiments indicated by the black circles in the bar graphs. Statistical significance determined by one-way ANOVA (ns = not significant, **p < 0.01, ***p < 0.001, ****p < 0.0001). (j) Flow cytometry plots of RPE FAP-TRF1 cells showing gating based on EdU and propidium iodine staining for various cell cycle phases 24 h after no treatment or exposure to dye, light, 5’ dye + light, 20 J/m² UVC, or one hour treatment with 2.5 or 10 mM KBrO₃. (k) Representative images of β-galactosidase positive BJ FAP-TRF1 cells obtained 4 days after recovery from indicated treatments (From Fig. 1h-i). Scale bar = 100 μm. (l) Size of nuclear area (μm²) of BJ (blue) or RPE (red) FAP-TRF1 cells obtained 4 days after recovery from no treatment or 5 min dye + light. Error bars represent the mean ± s.d. from the indicated n number of nuclei analyzed. Statistical analysis by two-tailed t-test (***p < 0.001, ****p < 0.0001).

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