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. 2019 Nov 19;10(1):231–253. doi: 10.1080/19491034.2019.1688932

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© 2019 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 5. — Induction of processive DNA synthesis repair leads to the decompaction of heterochromatin.

(a) Live-cell imaging plus 405 nm laser microirradiation of mCherry-PCNA expressing C2C12 cells. Constitutive heterochromatin was visualized by expressing GFP-tagged MeCP2. Full time lapse is shown in Movie 5. Enlarged regions represent sites of irradiation. Binary images: thresholded pictures of heterochromatic GFP-tagged MeCP2 and area size relative to pre-irradiation image. (b) Evaluation of MeCP2 marked area sizes at 60 s and 120 s after irradiation normalized to initial size are shown at the right (mean + standard error). n = between 19 and 22 cells per condition. (c) Recompaction of constitutive heterochromatin was visualized during a time course of 5 h. The decompaction of the heterochromatic compartment occurs within the first seconds to minutes. The compartment remains in the decompacted state for up to 2 h. Then, the recompaction starts as indicated by a decreasing area of the heterochromatic compartment. The recompaction is correlated with the release of PCNA from the microirradiated site (121 min). Scale bar 10 microns. (d) Mean areas of heterochromatic compartments are plotted over time after microirradiation with 405 nm laser and standard error is indicated by the shaded area. n = 10.