Fig. 2.

Association of the ERAD substrate glycoprotein to OS-9 requires EDEM1. HEK 293 cells expressing H2a, OS-9.1/2-S, XTP3-B-S, and anti-EDEM1 shRNA where indicated were lysed in mild conditions. Lysates (10% of total) were run on SDS-PAGE and immunoblotted with anti-S tag antibody or with anti-GAPDH (two bottom panels). The rest of the lysates were immunoprecipitated with anti-H2a, subjected to SDS-PAGE and immunoblotted with anti-S tag or anti-H2a (two upper panels). EDEM1 knockdown inhibited the association of the ERAD substrate with OS-9 by 80% and with XTP3-B by 25% (graph). The results shown are representative of three repeat experiments