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Figure 4


Fig. 4. Binding of BiP to luminal domain mutated Ire1. KMY1516 (ire1 null mutant) cells carrying pRS423-Ire1-HA (2-µm plasmid for expression of Ire1-HA) or mutant alleles were cultured with (+) or without (–) 2 µg/ml tunicamycin (Tun) for 60 minutes and their lysates were then used for anti-HA immunoprecipitation. The cell lysates (equivalent to 3x106 cells) and the anti-HA immunoprecipitates (equivalent to 1x107 cells; {alpha}-HA IP) were analyzed by anti-HA ({alpha}-HA) and anti-BiP ({alpha}-BiP) western blotting (WB). For the empty vector control ({Delta}ire1), pRS423 was used instead of pRS423-Ire1-HA.