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. 2020 Sep 25;11:4869. doi: 10.1038/s41467-020-18693-1

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© The Author(s) 2020

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. 2 — a Western blot analysis of fractionated extracts detects PARG-1 in all subcellular compartments with enrichment in the nuclear chromatin-bound fraction. GAPDH was used as a loading control of the cytosolic fraction and Histone H3 for the chromatin-bound fraction. Analysis was performed in biological duplicates. b Western blot analysis of fractionated extracts showing similar expression of GFP-tagged and untagged PARG-1. Analysis was performed in biological duplicates. c Top: PARG-1::GFP localization in a wild-type gonad. Scale bar 30 μm. Bottom: enlarged insets showing dynamic localization of PARG-1::GFP in different stages of meiotic prophase I. Scale bar 5 μm. Analysis was performed in biological triplicates. d Mid- and late-pachytene nuclei of parg-1::GFP co-stained for lateral (HTP-1 and -3) and central component (SYP-1) of the SC. Scale bar 5 μm. Analysis was performed in biological triplicates. e Late pachytene nuclei showing overlapping localization of PARG-1::GFP with OLLAS::COSA-1. Scale bar 5 μm. Analysis was performed in biological triplicates. f Late pachytene nuclei stained for HTP-1, SYP-1, and GFP showing localization of PARG-1 along chromosomes. In cosa-1 mutants, redistribution of PARG-1::GFP to the short arm of the bivalent is absent. Scale bar 5 μm. Analysis was performed in biological triplicates. g Impaired axes formation in htp-3 mutants prevents PARG-1::GFP localization. Arrow heads indicate regions of DNA devoid of PARG-1::GFP. Scale bar 5 μm. Analysis was performed in biological triplicates. h PARG-1::GFP associates with HTP-3 in late-pachytene nuclei in absence of synapsis. Scale bar 5 μm. Analysis was performed in biological duplicates. i Western blot analysis of endogenous PARG-1 on GFP pull downs performed in htp-3::GFP and GFP::syp-3 strains. Wild-type worms were used as the untagged negative control. Analysis was performed in biological duplicates. j Western blot analysis of endogenous PARG-1 on GFP pull downs performed in htp-1::GFP and rec-8:: GFP strains. Analysis was performed in biological duplicates.