Unravelling the sub-Golgi localization using En face and side-averaging

Abstract

Other than its two main functions being processing and transport, it is unknown how the Golgi complex works on a molecular or cellular level. The localization of proteins demonstrates their function in the Golgi. The lab has developed three methods used to study the localization of Golgi proteins. In this study, two methods, En face averaging and side-averaging were tested on twelve proteins. Super-resolution Airyscan microscopy was used to identify En face view and side view of individual nocodazole-induced Golgi ministacks. En face view demonstrates if the protein localizes to the interior or the rim, while side view shows the protein localization along the axis from the cis-Golgi to the trans-Golgi Network (TGN). The axial localization results generally corroborate with the previously published literature. The two methods used to study the localization of proteins provide unique insights into the molecular and cellular mechanisms governing the intra-Golgi transport.