Common Assays in Mammalian Golgi Studies

Abstract

The Golgi is a complex structure characterized by stacks of tightly aligned flat cisternae. In mammalian cells, Golgi stacks often concentrate in the perinuclear region and link together to form a ribbon. This structure is dynamic to accommodate continuous cargo flow in and out of the Golgi in both directions and undergoes morphological changes under physiological and pathological conditions. The fine, stacked Golgi structure makes it difficult to study by conventional light or even super-resolution microscopy. Furthermore, efforts to understand how Golgi structural dynamics impact cellular processes have been slow because of the knowledge gap in the protein machinery that maintains the complex and dynamic Golgi structure. In this method article, we list the common assays used in our research to help new and established researchers select the most appropriate method to properly address their questions.

Keywords: Fractionation; Function; Glycosylation; Golgi biology; In vitro techniques; Microscopy; Morphology; Proteomics; Proximity labeling; RNA interference; RNA-seq; Secretomics; Trafficking; Transfection; Viral system.