Palmitoylation-dependent probes for labeling the Golgi apparatus

Small-molecule fluorescent probes that selectively stain the Golgi apparatus in living cells are valuable tools for investigating Golgi-associated biology and disease. Conventional ceramide-based stains have been widely used, but they require complex staining procedures and often exhibit low Golgi specificity. To address these limitations, we recently developed a series of fluorescent probes incorporating the tri-N-methylated myristoyl-Gly-Cys (myrGC^3Me) motif, which targets the Golgi membrane via S-palmitoylation. Modular conjugation of this motif with organic fluorescent dyes yielded blue, green, and red fluorescent probes. Here, we provide detailed procedures for the chemical synthesis and application of these probes to enable simple and rapid staining of the Golgi in living cells with high specificity and negligible cytotoxicity. We also describe procedures for visualizing dynamic changes in Golgi morphology during drug treatments and cell division.