Mouse anti-Lamp1 mAb (H4A3; 1:500 for immunofluorescence or hereafter IF), mouse anti-CD63 mAb (H5C6; 1:100 for IF), and mouse anti-CD8a mAb (OKT8; 1:500 for IF) were from the Developmental Studies Hybridoma Bank. Mouse anti-GFP mAb (catalog no. sc-9996; 1:1000 for Western blotting) was purchased from Santa Cruz Biotechnology, Inc. Mouse anti-EEA1 mAb (catalog no. 610456; 1:500 for IF) and mouse anti-GM130 mAb (catalog no. 610823; 1:500 for IF) were from BD Biosciences. Horseradish peroxidase–conjugated goat anti-mouse (catalog no. 176516; 1:10,000 for Western blotting) was from Bio-Rad. Alexa Fluor–conjugated goat antibodies against mouse IgG (1:500 for IF) were from Thermo Fisher Scientific.
PNGase F was purchased from New England Biolabs. The following small molecules were commercially available: nocodazole (Merck; working concentration: 33 μm ), brefeldin A (Epicenter Technologies; working concentration: 10 µg/ml), cycloheximide (Sigma–Aldrich; working concentration: 10 µg/ml), and GalNAc-O-Bn (Sigma–Aldrich; working concentration: 2 mm ).
PNGase F was purchased from New England Biolabs. The following small molecules were commercially available: nocodazole (Merck; working concentration: 33 μ