Deoxymannojirimycin

Human HEK293T, MDA-MB-231, and mouse mammary carcinoma 4T1 cells, were bought from the American Type Culture Collection (Manassas, VA) and maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS, Gibico) at 37 °C under 5% CO₂. HCC1806 cells were maintained in RPMI-1640 medium supplemented with 10 % FBS at 37 °C under 5% CO₂. All the cells were authenticated using short-tandem repeat profiling, and tested negative for mycoplasma contamination. Compounds MG132 (S2619, Selleck Chemical), cycloheximide (BS168A, Biosharp), Thiamet G (HY-12588, MCE), PUGNAc (A7229, Sigma), tunicamycin (T7765, Sigma), 2F-Peracetyl-Fucose (344827, Sigma), swainsonine (16860, Cayman Chemical), and deoxymannojirimycin (D9169, Sigma) were obtained commercially.

A Cosmosil 5C18-AR column (0.60 × 25 cm) was purchased from NacalaiTesque, Inc. (Kyoto, Japan), and a Shodex Asahipak NH2P-50 column (0.46 × 25 cm) was purchased from Showa Denko Co. (Tokyo, Japan). Man_9-5GlcNAc₁-PA and Man_9-5GlcNAc₁-PA were prepared as described in previous reports (Kimura et al., 2000 (link); Kimura and Matsuo, 2000 (link)). α-1,2-Man’ase from Aspergillus saitoi was purchased from ProZyme, Inc. (Hayward, CA, United States). Swainsonine, deoxymannojirimycin, deoxynojirimycin, and Jack bean β-GlcNAc’ase were purchased from Sigma-Aldrich (St. Louis, MO, United States), and Endo-H was purchased from Promega (Madison, WI, United States). Glc3Man9GlcNAc2-PA and Glc2Man9GlcNAc2-PA were purchased from Masuda Chemical Industries Co. (Kagawa, Japan).