Figure S5. Characteristics of the CRISPR/Cas9-mediated IRE1B mutant lines #2–5 and #9–6.

(A) Genotyping of IRE1A-C genes in wild-type, ire1a ire1b, ire1a ire1c mutants, and T₃ plants of lines #2–5 and #9–6. Lane M, 100 bp DNA ladder. Lanes 1–6, six independent plants for each line. Note that PCR amplification of Cas9 gene was performed to detect T-DNA and that #9–6 lane 7 (asterisk) is a T₂ sibling plant having T-DNA used as a control. (B) RT-PCR of IRE1B mRNA in ire1a ire1c, ire1a ire1b mutants, and T₃ plants of #2–5 and #9–6. Actin2 (Act2) was used as an internal control. (C) DTT sensitivity of the ire1a ire1b, ire1a ire1c mutants, and T₃ plants of #2–5 and #9–6. Seedlings at 15 DAG were treated with or without 1 mM DTT. M, mutant; W, wild-type.