LMAN1 deletion mutants have been described previously (38 (link)) and were subcloned into pKH3 or pK-Myc vectors (46 (link)). Gateway vectors expressing humanized BirA* bearing R118G cDNA were kind gifts from A. Gingras, Lunenfeld-Tanenbaum Research Institute, Toronto (47 (link)). Both human SAR1A and SAR1B were placed in the N terminus of BirA*-FLAG, and SAR1B was primarily used in this study. The RUSH constructs (streptavidin-KDEL-IRES-SBP-mCherry-MAN II) were obtained from Addgene, and MAN II was replaced by LMAN1 cDNA. Other expression constructs have been described previously (48 (link)). All constructs were confirmed by complete DNA sequencing. Chemicals were purchased from Sigma-Aldrich.
The anti-SEC24A antibody has been previously described (48 (link)). Rabbit anti-FLAG antibody, mouse anti-actin antibody, and rabbit anti-SEC23 antibody were purchased from Sigma-Aldrich. Rabbit anti-LMAN1 antibodies were purchased from Stressgen. Preparation of anti-SAR1, anti-ribophorin, and anti-SEC22B antibodies has been described previously (49 (link)).
The anti-SEC24A antibody has been previously described (48 (link)). Rabbit anti-FLAG antibody, mouse anti-actin antibody, and rabbit anti-SEC23 antibody were purchased from Sigma-Aldrich. Rabbit anti-LMAN1 antibodies were purchased from Stressgen. Preparation of anti-SAR1, anti-ribophorin, and anti-SEC22B antibodies has been described previously (49 (link)).
