Figure 4: GRASP depletion enhances membrane association of coat proteins.
From: Regulation of protein glycosylation and sorting by the Golgi matrix proteins GRASP55/65
HeLa cells transfected with indicated RNAi were subjected to subcellular fractionation. Proteins in the cytosol and membrane fractions were analysed by western blot for β-COP (a), δ-COP (b), clathrin heavy chain (CHC, c), sec31 (d), bet1 (e) and α-tubulin (f). In some panels, three times of the membrane fractions were loaded onto the SDS–polyacrylamide gel electrophoresis (lane 3). Shown are representative images from three independent experiments. Quantification results (mean±s.e.m.) are shown on the right to indicate the percentage of the protein found in the membrane fractions. (g) GRASP depletion causes vesiculation of the Golgi. Quantitation of electron microscope images of GRASP-depleted cells for the number of vesicles adjacent to a Golgi stack. Shown are representative results (mean±s.e.m. from 20 different cells) from three independent experiments. Cells with both GRASP55 and GRASP65 depleted did not usually contain distinguishable Golgi stacks and thus were not quantified.
