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. 2022 Apr 30;50(9):5047–5063. doi: 10.1093/nar/gkac309

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© The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 4. — Telomeres are composed of a heterogeneous mesh of chromatin fibres. (A–E) Tomographic reconstructions of MEFs transfected with eGFP–APEX2–TRF1 fixed under isotonic conditions (A, B) or after hypotonic treatment (C–E). (A, C) From left to right: 0 degree projection image, higher magnification thereof, averaged tomographic slices of the same telomere with a thickness of 5.4 nm before (unfiltered) and after filtering to 2 nm. (B, D) Surface rendering of the same telomeres and slices shown in panels (A) and (C). Left: Full volume. Right: Slice with a thickness of 5.4 nm. Blue lines indicate the mesh of chromatin fibres. (E) Volume surface rendering of telomeric chromatin structures. Selected fibre diameters measured along the dashed lines are specified in nm. (F) Evaluation of chromatin fibre diameters of MEFs and U2OS cells transfected with eGFP–APEX2–TRF2 (TRF2), eGFP–APEX2–TRF1 (TRF1), eGFP–APEX2–H2B (H2B) or without APEX label from areas surrounding telomeres that are lightly UA stained only (UA only). Iso. = isotonic conditions. Hypo. = hypotonic treatment. Asterisks denote statistical significance based on two-sided t-tests: ***P < 0.001, **P < 0.01 and *P < 0.05. Median values (Ø) and number of cells analysed (n) for each condition are specified at the top of the graph. (G) Volume surface rendering of chromatin fibres of cells fixed under isotonic conditions. Labels equivalent to panel (F).