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ASCB | EMBO San Diego 2018
Annual Meeting of the American Society for Cell Biology and the European Molecular Biology Organization

Dec 08-12, 2018, San Diego, CA, USA
Booth Nr. 1137 (of 89Nort / Chroma Technology)

 

BPS Baltimore 2019
63rd Annual Meeting of the Biophysical Society

Mar 2-6, 2019, Baltimore, Maryland, USA
Booth Nr. 503

 

NWG Göttingen 2019
13th Meeting of the German Neuroscience Society

Mar 20-23, 2019, Göttingen, Germany
Booth Nr. 8

DNA Repair Pathways

Activation of multiple DNA Repair Pathways by Sub-nuclear Damage Induction Methods

  

The DNA damage in cultured living cells was induced by localized 266 nm radiation (sub-nuclear damage induction). The cells expressed GFP tagged repair factors GFP-XPA and Rad54-GFP to follow the nucleotide excision repair (NER) and double strand break repair (DSB) mechanisms, respectively.

 

Experimental information: 


Imaging: Zeiss Axiovert 200M microscope modified for UV transmission equipped with LSM510 confocal module

 

DNA damage: 2 mW 266 nm pulsed (7.8 kHz) DPSS laser (DPSL-266) and fixed spot illumination system (AiWon) (Rapp OptoElectronic GmbH)

 

 tl_files/rapp/images/spot and pattern illumination/DNA repair.jpg

 

(A) GFP-XPA expressing cells were irradiated with 266 nm either without (arrow) or with attenuation (arrowhead). GFP-XPA accumulates on both areas (green, left panel) whereas TUNEL (red, middle panel) only stains positive on the spot that was created without attenuation. (B) GFP-XPA expressing cells were irradiated by attenuated UV-C laser light (arrow). Presence of CPDs was shown by immunohistochemical staining with -CPD

(red, middle panel). (C) Cells that were irradiated in G1 or G2 phase (homogeneous PCNA pattern, red, middle panel) show no accumulation of Rad54-GFP (green, left panel) 2 hours after irradiation (arrow). (D) In cells that were irradiated in S phase (PCNA pattern in foci, red, middle panel) Rad54-GFP (green, left panel) accumulates at locally irradiated areas within 1 hour after irradiation (arrow).


Data from: Dinant et al. (2007) Activation of multiple DNA Repair Pathways by Sub-nuclear Damage Induction Methods, J. Cel. Sci. 120, 2731-40.

 

 
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