Accedi

Asan Medical Center

4 ARTICLES PUBLISHED IN JoVE

Genetics

RNA Interference-based Investigation of the Function of Heat Shock Protein 27 during Corneal Epithelial Wound Healing

Aeri Yoo^*1, Hyun-Min Park^*2, Soon-Suk Kang², Eun-Soon Kim², Hungwon Tchah², Jae Yong Kim²

¹Department of Ophthalmology, Saevit Eye Hospital, ²Department of Ophthalmology, University of Ulsan College of Medicine, Asan Medical Center

Herein, we present a protocol to use heat shock protein 27 (HSP27)-specific small interfering RNA to assess the function of HSP27 during corneal epithelial wound healing. RNA interference is the best method for effectively knocking-down gene expression to investigate protein function in various cell types.

Biology

Sample Preparation and Imaging of Exosomes by Transmission Electron Microscopy

Min Kyo Jung¹, Ji Young Mun²

¹Department of Convergence Medicine, University of Ulsan College of Medicine and Asan Institute for Life Sciences, Asan Medical Center, ²Synaptic Circuit Plasticity Laboratory, Department of Structure and Function of Neural Network, Korea Brain Research Institute

This protocol describes the various techniques necessary for transmission electron microscopy including negative staining, ultrathin sectioning for detailed structure, and immuno-gold labelling to determine the positions of specific proteins in exosomes.

Immunology and Infection

Immunostimulatory Agent Evaluation: Lymphoid Tissue Extraction and Injection Route-Dependent Dendritic Cell Activation

Jun-O Jin^1,2, Soyeong Jang³, Hyehyun Kim⁴, Junghwan Oh⁴, Sungbo Shim⁵, Minseok Kwak^3,4, Peter C.W. Lee⁶

¹Shanghai Public Health Clinical Center, Shanghai Medical College, Fudan University, ²Department of Medical Biotechnology, Yeungnam University, ³Department of Chemistry, Pukyong National University, ⁴Marine-integrated Bionics Research Center, Pukyong National University, ⁵Department of Biochemistry, Chungbuk National University, ⁶Department of Biomedical Sciences, University of Ulsan College of Medicine, Asan Medical Center

Experimental procedures for the subsequent extraction of lymphatic tissues to test lymphoid dendritic cell activation are described after treatment of an immunostimulating nanomaterial.

Biology

A Novel Nicotinamide Adenine Dinucleotide Correction Method for Intracellular Ca²⁺ Measurement with Fura-2-Analog in Live Cells

Jeong Hoon Lee¹, Jeong Mi Ha¹, Quynh Mai Ho¹, Chae Hun Leem^1,2,3

¹Department of Physiology, University of Ulsan College of Medicine/Asan Medical Center, ²Asan Medical Center, ³Asan Medical Institute of Convergence Science and Technology

Due to the spectral overlapping of the excitation and emission wavelengths of NADH and fura-2 analogs, the signal interference from both chemicals in live cells is unavoidable during quantitative measurement of [Ca²⁺]. Thus, a novel online correction method of NADH signal interference to measure [Ca²⁺] was developed.