Supplemental Figure 3

1. Supplemental Figure 3 Effects of b-GlcY and a-GalY on the expression of b-GlcY-inducible genes, GA-inducible genes and repressors of GA signaling. Real-time RT-PCR and semi-quantitative RT-PCR were performed using cDNA from 1.5 g total RNA, which were the same RNA samples as used in Fig. 5, isolated from 2.5 x 105 protoplasts. Aleurone protoplasts were treated with DMSO as a control or GA3 plus a-GalY or GA3 plus b-GlcY for 24 hours. (A) b-GlcY-inducible genes, (B) GA-inducible genes, (C) repressors of GA signaling. In real-time RT-PCR analysis, data shown are transcript levels relative to mock-treated samples (control = 100) and ubiquitin-conjugating enzyme (Contig4868_at) was used as an internal standard. In semi-quantitative RT-PCR analysis, GAPDH (Contig149_at) and PolyA-binding protein (Contig546_at) genes were chosen for internal controls. The primers used in this study are listed in Supplemental Table 5.

2. A +GA3 +GA3 Cont -GalY -GlcY Cont -GalY -GlcY Cont -GalY -GlcY Cont -GalY -GlcY Cont -GalY -GlcY Cont -GalY -GlcY Cont -GalY -GlcY Cont -GalY -GlcY Cont -GalY -GlcY +GA3 +GA3 +GA3 +GA3 +GA3 +GA3 +GA3 FAD-linked oxidoreductase Chitinase class I Chitinase III Blue copper-binding protein Oxalate oxidase Permatin PR5 GAPDH polyA-binding protein HvBWMK1 Ferritin Relative transcript level (transcript of control = 100) Pirin B Amy32b EPB1 Hypothetical protein GAPDH polyA-binding protein Relative transcript level (transcript of control = 100) C HvWRKY38 HvWRKY51 HvEsi47 Relative transcript level (transcript of control = 100)