Giant unilamellar vesicles (GUVs) are large, spherical lipid bilayer vesicles typically ranging from 1 to 100 micrometers in diameter, far exceeding the size of typical small unilamellar vesicles (SUVs) employed, for example, in liposome floatation assays. Their unique size and structural simplicity make them particularly suitable for a wide range of biophysical studies. GUVs serve as versatile model systems for studying membrane-protein interactions, membrane curvature sensing, and membrane remodeling processes. In addition, by micromanipulation, fluorescence recovery after photobleaching, and single-molecule imaging, membrane bending, deformation, and fusion events can be investigated with high spatial and temporal resolution. Here, we summarize our protocols generating GUVs for studying nucleoporin-membrane interaction including protocols reconstituting transmembrane nucleoporins into GUVs.