Figure 9.

Rot1 forms a ternary complex with BiP/Kar2 and Kre6. ROT1 and rot1-2 cells expressing HA-Kre6 (YM41 and YM42) were incubated for 10 min at 37°C, dissociated in PBS, and treated with 10 mM NaN₃ and 2 mM DSP (or the vehicle, DMSO only) for 1 h at 37°C. Double immunoprecipitation was performed against Rot1 first and then Kar2. Nonimmune serums (n.i.) were used for negative controls. The second immunoprecipitants were analyzed by Western blotting using anti-HA and anti-Kar2 antibodies.