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. 2005 Sep 26;391(Pt 1):135–142. doi: 10.1042/BJ20050640

Figure 2. UPRE–lacZ reporter assay to monitor in vivo activity of the Ire1 mutants.

Figure 2

As described under the Experimental section, IRE1 mutations were generated on the IRE1 plasmid pRS315-IRE1-HA by in vivo gap repair in the yeast Δire1 strain KMY1015 carrying the UPRE–lacZ reporter plasmid pCZY1. For the wild-type Ire1 (WT) and empty vector (Δire1) controls, cells were respectively transformed with non-linearized pRS315-IRE1-HA and pPR315. Then cultures were treated at 30 °C with 2 μg/ml tunicamycin or 3 mM DTT for 4 h, and subjected to assays for cellular β-galactosidase activity. Each value is the average±S.D. for three independent transformants and was normalized to the tunicamycin-treated wild-type Ire1 control, which was set at 100.