Fig. 3

Sec62 interacts with β-catenin. a DLD1 cells were transfected with Flag-Sec62. The whole cell extracts were immunoprecipitated with an anti-Flag antibody affinity resin. Sec62-binding proteins were resolved by SDS-PAGE, detected by silver staining and analyzed by mass spectrometry. b DLD1 cell lysates were immunoprecipitated with anti-Sec62 antibody or anti-β-catenin antibody. Immunoprecipitants were immunoblotted with the indicated antibodies. c DLD1 cells were transfected with the indicated vectors and immunofluorescence was performed using anti-β-catenin antibody. d GST pull-down assay was performed with purified His-Sec62 and GST-β-catenin or GST-GSK3β proteins. The amounts of GST fusion proteins used in the GST pull-down assay are shown in the lower panel. e GST pull-down assay was performed with GST-β-catenin and His-Sec62 (left). Schematic diagram of β-catenin deletion mutants (right). f GST pull-down assays was performed with GST-β-catenin and His-Sec62 (upper). A schematic diagram of β-catenin deletion mutants is shown in the lower panel. The numbers in blue box indicate the ARM repeats. g GST pull-down was performed with GST-Sec62 and His-β-catenin (upper). Shown is a schematic diagram of GST-Sec62 deletion mutants (lower). h GST pull-down assays were performed with GST-Sec62 or GST-Sec62 DA