Visualizing membrane contact site between the ER and Golgi to analyze the functional kinetics

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As a first step to visualize speculated membrane contact sites (MCS) between intracellular organelles such as the endoplasmic reticulum and Golgi apparatus, I proved the presence of the sites where these organelles membranes are closely apposed by Proximity Ligation Assay. Next, I tried to establish a split-DHFR system for real-time monitoring of MCS in vivo. At present, fairly good results are obtained and it is expected in near future to achieve the original goal of visualizing MCS in vivo by increasing the sensitivities and to clarify the mechanisms by which the transport of cholesterol is regulated.