Antibodies used for western blotting were as follows: rabbit polyclonal anti-Rer1 (R4407, Sigma-Aldrich, St. Louis, MO, USA), anti-Hrd1 (H7915, Sigma), anti-gp78 (9590, Cell Signaling Technology, Danvers, MA, USA), a rabbit monoclonal anti-Calnexin (C5C7, Cell Signaling), goat polyclonal anti-GFP (RDI, Fitzgerald Industry, Concord, MA, USA), mouse monoclonal anti-α-actin (C4, Millipore, Billerica, MA, USA), anti-FLAG (M2, Sigma-Aldrich), and anti-GSK3β (610201, BD Transduction Laboratories, San Diego, CA, USA) antibodies. Antibodies used for immunoprecipitation were as follows: mouse monoclonal anti-GFP (3E6, Q-Biogene, Carlsbad, CA, USA), anti-α-tubulin (DM1A, Sigma-Aldrich), and anti-FLAG (M2, Sigma-Aldrich) antibodies. Antibodies used for immunocytochemistry were as follows: rabbit polyclonal anti-Rab7 (a gift from Y. Wada, Osaka University)49 (link), anti-ERGIC-53 (E1031, Sigma-Aldrich), mouse monoclonal anti-PDI (1D3, Enzo Life Sciences, Plymouth Meeting, PA, USA), and anti-Lamp1 (H4A3, Santa Cruz Biotechnology, CA, USA) antibodies. Alexa Fluor® 555 or 594-conjugated anti-mouse or anti-rabbit immunoglobulin Gs (IgGs) (Life Technologies) were used as secondary antibodies.
MG132 was purchased from the Peptide Institute (Osaka, Japan). Bafilomycin A1 was purchased from Wako Pure Chemicals (Osaka, Japan). Cycloheximide was purchased from MP Biomedicals (Solon, OH, USA).
MG132 was purchased from the Peptide Institute (Osaka, Japan). Bafilomycin A1 was purchased from Wako Pure Chemicals (Osaka, Japan). Cycloheximide was purchased from MP Biomedicals (Solon, OH, USA).