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Figure 3


Fig. 3. HAC1u cleavage by the Ire1 luminal domain mutants. KMY1516 (ire1 null mutant) cells carrying pRS313-Ire1-HA (centromeric plasmid for expression of Ire1-HA) or mutant alleles were cultured with (+) or without (–) 2 µg/ml tunicamycin (Tun) for 60 minutes and total RNA was analyzed by northern blotting using the HAC1 probe. For the empty vector control ({Delta}ire1), pRS313 was used instead of pRS313-Ire1-HA. The positions of the uncleaved (HAC1u), the cleaved and ligated (HAC1i) and the cleaved but unligated (5'-cleaved and 3'-cleaved) versions of HAC1 mRNA are indicated (Kawahara et al., 1998). The percentage of HAC1 mRNA cleavage was calculated as described in the Materials and Methods.





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