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. 2023 Apr 14;14:2144. doi: 10.1038/s41467-023-37761-w

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© The Author(s) 2023, corrected publication 2023

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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Fig. 5 — a IF-FISH showing cells containing ≥4 UTs colocalized with S9.6 antibody (green) in Rap1^–/– MEFs cells treated with shTrf2 + TRF2^ΔB. Scale bars: 5 µm. b Quantification of percent UTs colocalized with S9.6 foci antibody in (a). Data represents the mean of three independent experiments ± SD from a minimum 300 nuclei analyzed per experiment. ****P < 0.0001 by one-way ANOVA. ns non-significant. c IF-FISH showing cells containing ≥4 UTs colocalized with TERRA (green) in Rap1^–/– MEFs cells treated with shTrf2 + TRF2^ΔB. Scale bars: 5 µm. d Quantification of percent UTs colocalized with TERRA in (c). Data represents the mean of three independent experiments ± SD from a minimum 350 nuclei analyzed per experiment. ***P = 0.0001, ****P < 0.0001 by one-way ANOVA. ns non-significant. e Quantification of UT frequencies after doxycycline induced RNaseH1^WT or catalytically dead RNaseH1^D210N mutant in U2OS cells expressing indicated constructs. Data represents the mean of three independent experiments ± SD from a minimum of 350 nuclei analyzed per experiment. *P = 0.0325, **P = 0.0035, ****P < 0.0001 by one-way ANOVA. ns: non-significant. f IP-MS data using Flag-TRF2 and Flag-vector as baits showing R-loop associated proteins coprecipitated with Flag-WT TRF2 but not in Flag-vector in one representative experiment. g Co-IP with Flag antibody with lysates from 293 T cells expressing indicated proteins showing the interaction between TRF2 and DDX21 mutant depends on the DDX21 C-terminus. The blot shown is the representative of two independent experiments. h Quantification of UT frequencies in U2OS cells expressing Flag-DDX21^WT and the indicated DDX21 deletion mutants in the presence of shTRF2 + TRF2^ΔB;L288R. Data represents the mean of three independent experiments ± SD from a minimum 300 nuclei analyzed per experiment. *P = 0.0254, **P = 0.0064 by one-way ANOVA. ns non-significant. i Quantification of UT frequency upon overexpression of GFP ADAR1p110 and catalytically inactive ADAR1p110^E912A mutant in U2OS cells expressing indicated DNA constructs. Data represents the mean of three independent experiments ± SD from a minimum 350 nuclei analyzed per experiment. ***P = 0.0001, ****P < 0.0001 by one-way ANOVA. ns non-significant. Source data are provided as a Source Data file.