L-2HG and LDHC guarded centromere morphology in diakinesis cells.

<p>(A) Experimental Design (Created with Biorender). This experiment aimed to study the effect of 2HG on compacting oxamate-decompacted diakinesis centromeres. Diakinesis cells were isolated from two groups: mice injected once with oxamate (1.3 g/kg) and sacrificed after 24 hours (Ox) and mice...

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1. autor: Nina Mayorek (241888) (author)
Kolejni autorzy: Miriam Schlossberg (21696644) (author), Yousef Mansour (21696647) (author), Nir Pillar (5249612) (author), Ilan Stein (131429) (author), Fatima Mushasha (21696650) (author), Guy Baziza Paz (21696653) (author), Eleonora Medvedev (21696656) (author), Zakhariya Manevitch (21696659) (author), Julia Menzel (12879920) (author), Elina Aizenshtein (21696662) (author), Boris Sarvin (8232306) (author), Nikita Sarvin (21696665) (author), Erwin Goldberg (2868524) (author), Bryan A. Niedenberger (21696668) (author), Christopher B. Geyer (13840432) (author), Tomer Shlomi (22999) (author), Michael Klutstein (22329) (author), Eli Pikarsky (103960) (author)
Wydane: 2025
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_version_ 1851484667174191104
author Nina Mayorek (241888)
author2 Miriam Schlossberg (21696644)
Yousef Mansour (21696647)
Nir Pillar (5249612)
Ilan Stein (131429)
Fatima Mushasha (21696650)
Guy Baziza Paz (21696653)
Eleonora Medvedev (21696656)
Zakhariya Manevitch (21696659)
Julia Menzel (12879920)
Elina Aizenshtein (21696662)
Boris Sarvin (8232306)
Nikita Sarvin (21696665)
Erwin Goldberg (2868524)
Bryan A. Niedenberger (21696668)
Christopher B. Geyer (13840432)
Tomer Shlomi (22999)
Michael Klutstein (22329)
Eli Pikarsky (103960)
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author_facet Nina Mayorek (241888)
Miriam Schlossberg (21696644)
Yousef Mansour (21696647)
Nir Pillar (5249612)
Ilan Stein (131429)
Fatima Mushasha (21696650)
Guy Baziza Paz (21696653)
Eleonora Medvedev (21696656)
Zakhariya Manevitch (21696659)
Julia Menzel (12879920)
Elina Aizenshtein (21696662)
Boris Sarvin (8232306)
Nikita Sarvin (21696665)
Erwin Goldberg (2868524)
Bryan A. Niedenberger (21696668)
Christopher B. Geyer (13840432)
Tomer Shlomi (22999)
Michael Klutstein (22329)
Eli Pikarsky (103960)
author_role author
dc.creator.none.fl_str_mv Nina Mayorek (241888)
Miriam Schlossberg (21696644)
Yousef Mansour (21696647)
Nir Pillar (5249612)
Ilan Stein (131429)
Fatima Mushasha (21696650)
Guy Baziza Paz (21696653)
Eleonora Medvedev (21696656)
Zakhariya Manevitch (21696659)
Julia Menzel (12879920)
Elina Aizenshtein (21696662)
Boris Sarvin (8232306)
Nikita Sarvin (21696665)
Erwin Goldberg (2868524)
Bryan A. Niedenberger (21696668)
Christopher B. Geyer (13840432)
Tomer Shlomi (22999)
Michael Klutstein (22329)
Eli Pikarsky (103960)
dc.date.none.fl_str_mv 2025-07-10T17:56:42Z
dc.identifier.none.fl_str_mv 10.1371/journal.pgen.1011785.g005
dc.relation.none.fl_str_mv https://figshare.com/articles/figure/L-2HG_and_LDHC_guarded_centromere_morphology_in_diakinesis_cells_/29537597
dc.rights.none.fl_str_mv CC BY 4.0
info:eu-repo/semantics/openAccess
dc.subject.none.fl_str_mv Cell Biology
Genetics
Molecular Biology
Developmental Biology
Biological Sciences not elsewhere classified
spindle assembly checkpoint
satellite rna expression
sac ), suggesting
producing functional spermatozoa
histone methylation changes
including chromocenter clustering
2hg ), yet
regulates centromere condensation
hydroxyglutarate regulates centromere
chromocenter condensation
centromere dysfunction
vivo results
synaptonemal complex
localizing along
heterochromatin conformation
epigenetic modifications
diplotene stages
acute depletion
dc.title.none.fl_str_mv L-2HG and LDHC guarded centromere morphology in diakinesis cells.
dc.type.none.fl_str_mv Image
Figure
info:eu-repo/semantics/publishedVersion
image
description <p>(A) Experimental Design (Created with Biorender). This experiment aimed to study the effect of 2HG on compacting oxamate-decompacted diakinesis centromeres. Diakinesis cells were isolated from two groups: mice injected once with oxamate (1.3 g/kg) and sacrificed after 24 hours (Ox) and mice injected with vehicle-only (Ctr). Diakinesis cells from oxamate-injected mice, displaying enlarged centromeres (see B), were subjected to a 10-minute incubation at 32°C with 0.6 mM octyl L/D-2HG, while cells from control mice were treated under identical conditions with vehicle only. After incubation, nuclear spreads were prepared and immunostained for various markers: CREST/SCP3 (B-D, K), LDHC/SCP3 (H-J), CENP-C (E), CENP-N (F), and CENP-A (G). Representative images of E-G are presented in (<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1011785#pgen.1011785.s005" target="_blank">S5 Fig</a>). Cross-sectional area measurements of centromeres immunostained with the above antibodies were conducted (B-C, E-H). Another group of diakinesis cells underwent the same procedure but were incubated with 5 mM lactate instead of 2HG, and centromeres were immunostained with CREST antibodies (N). Each point in (B-C, E-H, and N) represents the mean area of all centromeres in one cell, while in (L-M), each point represents the mean staining intensity of centromeres in one cell. Data are presented as mean ± SE of n = 30-60 cells in each treatment. Statistical analyses were conducted using t-test for (B) and Tukey’s multiple comparisons test for (C-H and L-N). Immunostained areas were measured using Zeiss ZEN 3.3 software and expressed as a fraction of the mean of all experimental groups in arbitrary units (a.u). (D and I) Confocal images of centromere areas immunostained with CREST antibodies (quantified in C) and LDHC (quantified in H) are depicted. Scale bars in D and I = 10 µm. (J-K) High-resolution images of centromeres captured by the N-SIM microscope are provided, scale bar = 1 µm.</p>
eu_rights_str_mv openAccess
id Manara_20d06676b51c81d7533bd8f17c9c77d4
identifier_str_mv 10.1371/journal.pgen.1011785.g005
network_acronym_str Manara
network_name_str ManaraRepo
oai_identifier_str oai:figshare.com:article/29537597
publishDate 2025
repository.mail.fl_str_mv
repository.name.fl_str_mv
repository_id_str
rights_invalid_str_mv CC BY 4.0
spelling L-2HG and LDHC guarded centromere morphology in diakinesis cells.Nina Mayorek (241888)Miriam Schlossberg (21696644)Yousef Mansour (21696647)Nir Pillar (5249612)Ilan Stein (131429)Fatima Mushasha (21696650)Guy Baziza Paz (21696653)Eleonora Medvedev (21696656)Zakhariya Manevitch (21696659)Julia Menzel (12879920)Elina Aizenshtein (21696662)Boris Sarvin (8232306)Nikita Sarvin (21696665)Erwin Goldberg (2868524)Bryan A. Niedenberger (21696668)Christopher B. Geyer (13840432)Tomer Shlomi (22999)Michael Klutstein (22329)Eli Pikarsky (103960)Cell BiologyGeneticsMolecular BiologyDevelopmental BiologyBiological Sciences not elsewhere classifiedspindle assembly checkpointsatellite rna expressionsac ), suggestingproducing functional spermatozoahistone methylation changesincluding chromocenter clustering2hg ), yetregulates centromere condensationhydroxyglutarate regulates centromerechromocenter condensationcentromere dysfunctionvivo resultssynaptonemal complexlocalizing alongheterochromatin conformationepigenetic modificationsdiplotene stagesacute depletion<p>(A) Experimental Design (Created with Biorender). This experiment aimed to study the effect of 2HG on compacting oxamate-decompacted diakinesis centromeres. Diakinesis cells were isolated from two groups: mice injected once with oxamate (1.3 g/kg) and sacrificed after 24 hours (Ox) and mice injected with vehicle-only (Ctr). Diakinesis cells from oxamate-injected mice, displaying enlarged centromeres (see B), were subjected to a 10-minute incubation at 32°C with 0.6 mM octyl L/D-2HG, while cells from control mice were treated under identical conditions with vehicle only. After incubation, nuclear spreads were prepared and immunostained for various markers: CREST/SCP3 (B-D, K), LDHC/SCP3 (H-J), CENP-C (E), CENP-N (F), and CENP-A (G). Representative images of E-G are presented in (<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1011785#pgen.1011785.s005" target="_blank">S5 Fig</a>). Cross-sectional area measurements of centromeres immunostained with the above antibodies were conducted (B-C, E-H). Another group of diakinesis cells underwent the same procedure but were incubated with 5 mM lactate instead of 2HG, and centromeres were immunostained with CREST antibodies (N). Each point in (B-C, E-H, and N) represents the mean area of all centromeres in one cell, while in (L-M), each point represents the mean staining intensity of centromeres in one cell. Data are presented as mean ± SE of n = 30-60 cells in each treatment. Statistical analyses were conducted using t-test for (B) and Tukey’s multiple comparisons test for (C-H and L-N). Immunostained areas were measured using Zeiss ZEN 3.3 software and expressed as a fraction of the mean of all experimental groups in arbitrary units (a.u). (D and I) Confocal images of centromere areas immunostained with CREST antibodies (quantified in C) and LDHC (quantified in H) are depicted. Scale bars in D and I = 10 µm. (J-K) High-resolution images of centromeres captured by the N-SIM microscope are provided, scale bar = 1 µm.</p>2025-07-10T17:56:42ZImageFigureinfo:eu-repo/semantics/publishedVersionimage10.1371/journal.pgen.1011785.g005https://figshare.com/articles/figure/L-2HG_and_LDHC_guarded_centromere_morphology_in_diakinesis_cells_/29537597CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/295375972025-07-10T17:56:42Z
spellingShingle L-2HG and LDHC guarded centromere morphology in diakinesis cells.
Nina Mayorek (241888)
Cell Biology
Genetics
Molecular Biology
Developmental Biology
Biological Sciences not elsewhere classified
spindle assembly checkpoint
satellite rna expression
sac ), suggesting
producing functional spermatozoa
histone methylation changes
including chromocenter clustering
2hg ), yet
regulates centromere condensation
hydroxyglutarate regulates centromere
chromocenter condensation
centromere dysfunction
vivo results
synaptonemal complex
localizing along
heterochromatin conformation
epigenetic modifications
diplotene stages
acute depletion
status_str publishedVersion
title L-2HG and LDHC guarded centromere morphology in diakinesis cells.
title_full L-2HG and LDHC guarded centromere morphology in diakinesis cells.
title_fullStr L-2HG and LDHC guarded centromere morphology in diakinesis cells.
title_full_unstemmed L-2HG and LDHC guarded centromere morphology in diakinesis cells.
title_short L-2HG and LDHC guarded centromere morphology in diakinesis cells.
title_sort L-2HG and LDHC guarded centromere morphology in diakinesis cells.
topic Cell Biology
Genetics
Molecular Biology
Developmental Biology
Biological Sciences not elsewhere classified
spindle assembly checkpoint
satellite rna expression
sac ), suggesting
producing functional spermatozoa
histone methylation changes
including chromocenter clustering
2hg ), yet
regulates centromere condensation
hydroxyglutarate regulates centromere
chromocenter condensation
centromere dysfunction
vivo results
synaptonemal complex
localizing along
heterochromatin conformation
epigenetic modifications
diplotene stages
acute depletion