Nanjing University 8 articles published in JoVE Developmental Biology Generation of a Mouse Artificial Decidualization Model with Ovariectomy for Endometrial Decidualization Research Yang Zhang*1,2,3, Zhe Zhang*1,2,3, Nannan Kang1,2,3, Xiaoqiang Sheng1,2,3 1Center for Reproductive Medicine and Obstetrics and Gynecology, The Affiliated Drum Tower Hospital of Nanjing University Medical School, 2State Key Laboratory of Reproductive Medicine, Nanjing Medical University, 3Center for Molecular Reproductive Medicine, Nanjing University Here, we describe the method of generating an artificial decidualization model using the ovariectomized mouse, a classic endometrial decidualization experiment in the research field of endometrial decidualization. Biology 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. Biochemistry OaAEP1-Mediated Enzymatic Synthesis and Immobilization of Polymerized Protein for Single-Molecule Force Spectroscopy Yibing Deng1, Bin Zheng1, Yutong Liu1, Shengchao Shi1, Jingyuan Nie1, Tao Wu1, Peng Zheng1 1State Key Laboratory of Coordination Chemistry, School of Chemistry and Chemical Engineering, Nanjing University Here, we present a protocol to conjugate protein monomer by enzymes forming protein polymer with a controlled sequence and immobilize it on the surface for single-molecule force spectroscopy studies. Chemistry Stable DNA Motifs, 1D and 2D Nanostructures Constructed from Small Circular DNA Molecules Xin Guo1, Xue-Mei Wang1, Shou-Jun Xiao1 1State Key Laboratory of Coordination Chemistry, School of Chemistry and Chemical Engineering, Nanjing University This article presents a detailed protocol for T4 ligation and denaturing PAGE purification of small circular DNA molecules, annealing and native PAGE analysis of circular tiles, assembling and AFM imaging of 1D and 2D DNA nanostructures, as well as agarose gel electrophoresis and centrifugation purification of finite DNA nanostructures. Medicine Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography Jie Sun1, Guang Ming Yang2, Tao Tao1, Li Sha Wei1, Yang Pan2, Min Sheng Zhu1 1Model Animal Research Center and MOE Key Laboratory of Model Animal and Disease Study, Nanjing University, 2School of Pharmacy, Nanjing University of Chinese Medicine The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle. Biology Intracavernosal Pressure Recording to Evaluate Erectile Function in Rodents Feng Pan*2, Jie Zhang*3, Yuyan Liu1, Liangsheng Lu1, Xuefeng Qiu4, Kangtai Lv5, Qipeng Zhang1 1State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Jiangsu Engineering Research Center for microRNA Biology and Biotechnology, Nanjing University, 2Department of Andrology, The Affiliated Obstetrics and Gynecology Hospital of Nanjing Medical University, Nanjing Maternity and Child Health Care Hospital, 3School of Pharmaceutical Sciences, Nanjing Tech University, 4Department of Urology, Affiliated Drum Tower Hospital, School of Medicine, Nanjing University, 5Department of Ultrasound, Nanjing Qixia District Maternity and Child Health Care Hospital Intracavernosal pressure recording (ICP) is an important method to evaluate the erectile function of experimental animals. Here, a detailed protocol is demonstrated for the recording procedure of ICP by catheterizing the crura penis and then electrically stimulating the cavernous nerves in rats. Medicine Construction and Evaluation of a Murine Calvarial Osteolysis Model by Exposure to CoCrMo Particles in Aseptic Loosening Hui Jiang*1, Yicun Wang*1, Zhantao Deng*1,2,3, Jiewen Jin2,3, Jia Meng1, Shuo Chen1, Jun Wang1, Yang Qiu1, Ting Guo1, Jianning Zhao1 1Department of Orthopedics, Jinling Hospital, School of Medicine, Nanjing University, 2Center for Translational Medicine, Nanjing University Medical School, 3Jiangsu Key Laboratory for Molecular Medicine, Nanjing University Medical School This manuscript describes a murine calvarial osteolysis model by exposure to CoCrMo particles, which constitutes an ideal animal model for assessing the interactions between wear particles and various cells in aseptic loosening. Chemistry Fabrication of Spherical and Worm-shaped Micellar Nanocrystals by Combining Electrospray, Self-assembly, and Solvent-based Structure Control Xinyi Ding1,2,3, Yuxiang Sun1,2,3, Yanming Chen1,2,3, Wanchuan Ding1,2,3, Steven Emory1,2,3,4, Tianhao Li1,2,3, Zixing Xu1,2,3, Ning Han1,2,3, Jun Wang1,2,3, Gang Ruan1,2,3 1Department of Biomedical Engineering, College of Engineering and Applied Sciences, Nanjing University, 2Institute of Materials Engineering, College of Engineering and Applied Sciences, Nanjing University, 3Collaborative Innovation Center of Chemistry for Life Sciences, Nanjing University, 4Department of Chemistry, Western Washington University The present work describes a method to fabricate micellar nanocrystals, an emerging major class of nanobiomaterials. This method combines top-down electrospray, bottom-up self-assembly, and solvent-based structure control. The fabrication method is largely continuous, can produce high quality products, and possesses an inexpensive means of structure control.