Extended Data Fig. 8. Transactivation and binding of JAM2 to the vicinity of the transcription start site within the RD29B promoter.
a, Enhanced ABA response by ectopic expression of JAM2. Protoplasts of wild type were transfected with JAM2 effector cassette (3 µg DNA/105 protoplasts) and the promoter pRD29B or pRD29B with mutated ABA-responsive cis elements (ABRE), mABRE1 or mABRE2, driving luciferase expression. After incubation in the presence of 10 µM ABA for 16 h, the luciferase activity was measured (kRLU/RFU), (mean ± SD, n = 3). b, JAM2 interaction with RD29B is stimulated in the presence of ABA. Top, ChIP analysis of the promoter region of RD29B. Bottom, Scheme of pRD29B with the transcriptional start site at position 1. ABREs (motif ACGTG) are signified by filled boxes and their nucleotide distance relative to the start site is given with ABRE1 and ABRE2 closest and next closest to the start site respectively. Potential JAM2 binding sites, that is imperfect G-boxes with one nucleotide deviation from the core CACGTG motif83, are indicated with open boxes. Middle, the positions of the amplified DNA fragments are shown. Protoplasts were analysed after transient expression of JAM2 effector (3 µg DNA/105 protoplasts) or not (control) with or without 10 µM ABA (mean ± s.e.m., n = 3-4).