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Medical School of Nanjing University

3 ARTICLES PUBLISHED IN JoVE

Biology

Comet Assay as an Indirect Measure of Systemic Oxidative Stress

Lei Fang^1,2, Albert Neutzner^1,2, Stephanie Turtschi¹, Josef Flammer¹, Maneli Mozaffarieh¹

¹Department of Ophthalmology, University of Basel, ²Department of Biomedicine, University of Basel

Comet assay measures DNA breaks, induced by different factors. If all factors (except oxidative stress) causing DNA damage are kept constant, the amount of DNA damage is a good indirect parameter of oxidative stress. The goal of this protocol is to use comet assay for indirect measurement of oxidative stress.

Medicine

The Rodent Model of Nonarteritic Anterior Ischemic Optic Neuropathy (rNAION)

Yan Guo¹, Zara Mehrabian¹, Steven L. Bernstein^1,2

¹Department of Ophthalmology and Visual Sciences, University of Maryland-Baltimore School of Medicine, ²Anatomy and Neurobiology, University of Maryland-Baltimore School of Medicine

The following report describes how to replicate the rodent model of nonarteritic anterior ischemic optic neuropathy (rNAION), using the appropriate dye, contact lens and laser parameters. We also reveal the appropriate steps for evaluating the rNAION lesion in vivo.

JoVE Core

Mass Spectrometry Analysis to Identify Ubiquitylation of EYFP-tagged CENP-A (EYFP-CENP-A)

Yohei Niikura^*1, Lei Fang^*2, Risa Kitagawa^*3, Peizhao Li¹, Yao Xi¹, Ju You¹, Yan Guo², Katsumi Kitagawa³

¹MOE Key Laboratory of Model Animal for Disease Study, Model Animal Research Center, Nanjing University, ²Jiangsu Key Laboratory of Molecular Medicine, Medical School of Nanjing University, ³Greehey Children's Cancer Institute, Department of Molecular Medicine, UT Health Science Center San Antonio

CENP-A ubiquitylation is an important requirement for CENP-A deposition at the centromere, inherited through dimerization between cell division, and indispensable to cell viability. Here we describe mass spectrometry analysis to identify ubiquitylation of EYFP-tagged CENP-A (EYFP-CENP-A) protein.