Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2019 Dec 13;10:5693. doi: 10.1038/s41467-019-13674-5

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2019

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/.

PMC Copyright notice

Fig. 4 — a Determination of the minimal time permitting clearing of the FHIT gene from ongoing transcription. Upper panel: scheme of the experiments. Untreated cells (NT) and cells treated with 1 μM triptolide (Tpl) for the indicated periods of time were pulse labelled with EU for 30 min before recovery. Nascent RNAs were prepared by the Click-It method and quantified by RT-qPCR. Middle panel: map of the human FHIT gene with the position of exons (e1–e10), T-SDW/SDR and intronic primer pairs (i1–i8b) used for quantification. Lower panel: quantification of nascent RNA along FHIT at the different times of Tpl treatment. b Examples of chromosome breaks in cells treated with 600 nM Aph for 7 h. Left panel: metaphase plate displaying a break (red arrow) on Giemsa-stained chromosomes. Right panel: break at FRA3B (yellow arrow) visualized after DAPI staining and FISH with probes specific to the FHIT gene (green) and to the centromere of chromosome 3 (red). Contrast of DAPI-stained chromosomes (rightmost panel) was enhanced to better show the break. c Upper panel: scheme of the experiments: cells were treated with 1 μM triptolide and 600 nM Aph, alone or in combination, for the indicated times before metaphase preparation. In all conditions, cells were pulse labelled for 1 h with 30 µM BrdU at the beginning of the experiments and treated for 2 h with 200 nM nocodazole at the end of experiments to enrich the populations in metaphase cells. Lower panels: the fraction of BrdU-labelled metaphases was scored after DAPI staining and immunofluorescence revelation with anti-BrdU antibodies. Examples of BrdU-labelled and unlabelled metaphases are shown. Note also the presence of labelled and unlabelled interphase nuclei. d Determination of the frequencies of total chromosome breaks (left panel) and breaks at FRA3B (right panel). Breaks were scored as in b. Experiments shown in a, c and d were carried out twice and the error bars represent the SD. Source data are provided as a Source Data file.