Vacuoles were isolated from wild-type and vps35Δ S. cerevisiae cells grown to stationary phase. Peptides were digested using Preomics iST kit, analyzed by Thermo Orbitrap Eclipse in DDA mode with FAIMS (-45V/-65V). Data were searched against UniProt yeast database using Proteome Discoverer 2.4.1.15 with 10 ppm precursor and 0.02 Da fragment tolerances, filtered at 1% FDR. This dataset was used to identify retromer cargoes accumulating in vps35Δ vacuoles.