Perforated MDCK cells support intracellular transport

摘要: We have developed a method for perforating the plasma membrane of MDCK cells while retaining cellular functions. A nitrocellulose acetate filter was applied to apical side cells, grown on glass coverslip, and allowed dry. Segments adhered were detached from cell layer by shearing when peeled off. This macromolecules such as antibodies enzymes diffuse into cells. The otherwise intact judged light electron microscopy. perforated maintained their capacity support vesicular transport proteins lipids. Vesicular stomatitis virus infected readily incorporated [35S]methionine G protein following permeabilization. core-glycosylated during assembly in endoplasmic reticulum, further transported trans Golgi with high efficiency. Experiments using lipid probes demonstrated that newly synthesized fluorescent sphingolipids complex basolateral surface Our results show provide convenient efficient alternative cell-free assays studying molecular mechanism intracellular transport.