Figure 3
Induction of bZIP60 mRNA and activation of bZIP60 protein in the wild-type and ire1 mutants.
a, RNA blot analysis of bZIP60 in the wild-type and ire1a/ire1b treated with tunicamycin. RNA was prepared from seedlings treated with tunicamycin (5 mg/l) for the indicated period. b, Immunoblot analysis of bZIP60 in the wild-type and ire1a/ire1b. Protein extracts were prepared from seedlings treated as in a and immunoblot analysis was performed using anti-bZIP60 antibody. Ten micrograms of total protein was loaded in each lane. c, Immunoblot analysis of bZIP60 in the wild-type, bzip60, ire1a, ire1b and ire1a/ire1b. Seedlings treated with tunicamycin (5 mg/l) or DMSO (0.1%) as a control for 5 h were subjected to immunoblot analysis as in b. d, Immunoblot analysis of bZIP60 in the wild-type and in ire1a/ire1b by tunicamycin and DTT treatments. Seedlings were treated with DMSO (0.1%; -), tunicamycin (5 mg/l; Tm) and DTT (2 mM) for 5 h and used for protein extraction. bZIP60ΔC was transiently expressed in protoplasts prepared from bzip60 mutants and used for immunoblot analysis alongside. Positions of bZIP60u (u), bZIP60s (s) and bZIP60ΔC (ΔC) are indicated.
