High-throughput Evaluation of Protein Migration and Localization after Laser Micro-Irradiation.
DNA Breaks, Double-Stranded
/ radiation effects
DNA Damage
/ genetics
DNA Repair
/ genetics
Histone Code
/ genetics
Histones
/ chemistry
Humans
Lasers
/ adverse effects
Microscopy, Fluorescence
Phosphorylation
/ radiation effects
Protein Processing, Post-Translational
/ genetics
Proteins
/ chemistry
Single-Cell Analysis
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
28 02 2019
28 02 2019
Historique:
received:
12
11
2018
accepted:
01
02
2019
entrez:
1
3
2019
pubmed:
1
3
2019
medline:
17
9
2020
Statut:
epublish
Résumé
DNA- and histone-related research frequently comprises the quantitative analysis of protein modifications, such as histone phosphorylation. Analysis of accumulation and disappearance of protein foci are used to monitor DNA damage and repair kinetics. If the protein of interest doesn't accumulate in foci, laser micro-irradiation of single nuclei provides an alternative method to monitor DNA repair proteins and histone dynamics at the DNA damage site. We have developed an automated evaluation tool for standardized, high-throughput analysis of micro-irradiated cells featuring single cell background subtraction and detection across multiple fluorescence channels, allowing for robust statistics.
Identifiants
pubmed: 30816253
doi: 10.1038/s41598-019-39760-8
pii: 10.1038/s41598-019-39760-8
pmc: PMC6395640
doi:
Substances chimiques
Histones
0
Proteins
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
3148Subventions
Organisme : NIEHS NIH HHS
ID : R01 ES005775
Pays : United States
Organisme : NIGMS NIH HHS
ID : T32 GM007324
Pays : United States
Organisme : NCI NIH HHS
ID : R35 CA197574
Pays : United States
Organisme : NIDDK NIH HHS
ID : P01 DK057751
Pays : United States
Références
Stem Cell Reports. 2016 Dec 13;7(6):1013-1022
pubmed: 27974220
Oncogene. 2016 Feb 11;35(6):761-70
pubmed: 25961912
Methods Mol Biol. 2017;1599:277-285
pubmed: 28477126
J Vis Exp. 2018 Jan 31;(131):
pubmed: 29443023
Biol Cell. 2015 Dec;107(12):440-54
pubmed: 26482424
Int J Radiat Biol. 2018 Aug;94(8):708-718
pubmed: 29309203
Cold Spring Harb Symp Quant Biol. 1974;38:165-74
pubmed: 4133983
Nucleic Acids Res. 2016 Nov 2;44(19):9266-9278
pubmed: 27458206
J Cell Biol. 2010 May 31;189(5):777-82
pubmed: 20513764
J Vis Exp. 2017 Sep 5;(127):
pubmed: 28930988
Sci Rep. 2016 Jan 18;6:19567
pubmed: 26777522
Nucleic Acids Res. 2008 Oct;36(17):5678-94
pubmed: 18772227
Methods Mol Biol. 2017;1608:165-183
pubmed: 28695510
Radiat Res. 2017 Jul;188(1):114-120
pubmed: 28492345
Radiat Oncol. 2015 Aug 04;10:163
pubmed: 26238507
Indian J Cancer. 2004 Jan-Mar;41(1):47
pubmed: 15105580
Exp Cell Res. 1967 Feb;45(2):361-73
pubmed: 4164421
Int J Radiat Oncol Biol Phys. 2004 Feb 1;58(2):331-5
pubmed: 14751500
Acta Biol Med Ger. 1979;38(9):1285-93
pubmed: 399147