Supplemental Figure 3 Effects of -GlcY and -GalY on the expression of -GlcY-i nducible genes, GA-inducible genes and repressors of GA signaling. R eal-time RT-PCR and semi-quantitative RT-PCR were performed using cDN A from 1.5 g total RNA, which were the same RNA samples as used in F ig. 5, isolated from 2.5 x 10 5 protoplasts. Aleurone protoplasts wer e treated with DMSO as a control or GA 3 plus -GalY or GA 3 plus -GlcY for 24 hours. (A) -GlcY-inducible genes, (B) GA-inducible genes, (C) repressors of GA signaling. In real-time RT-PCR analysis, data s hown are transcript levels relative to mock-treated samples (control = 100) and ubiquitin-conjugating enzyme (Contig4868_at) was used as an inte rnal standard. In semi-quantitative RT-PCR analysis, GAPDH (Contig1 49_at) and PolyA-binding protein (Contig546_at) genes were chosen for inte rnal controls. The primers used in this study are listed in Suppleme ntal Table 5.
Supplemental Figure 3
Jan 26, 2016
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Supplemental Figure 3
Effects of -GlcY and -GalY on the expression of -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 -GalY or GA3 plus -GlcY for 24 hours. (A) -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.
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24 Cont 24 GA+alpha 24 GA+beta
Chitinase III
Blue copper-binding protein
Oxalate oxidase
Permatin PR5
GAPDH
polyA-binding protein
A
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1000
24 Cont 24 GA+alpha 24 GA+beta
Ferritin
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24 Cont 24 GA+alpha 24 GA+beta
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24 Cont 24 GA+alpha 24 GA+beta
Amy32b
Pirin
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80
120
160
24 Cont 24 GA+alpha 24 GA+beta
HvWRKY38
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160
24 Cont 24 GA+alpha 24 GA+beta
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24 Cont 24 GA+alpha 24 GA+beta
FAD-linked oxidoreductase
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24 Cont 24 GA+alpha 24 GA+beta
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24 Cont 24 GA+alpha 24 GA+beta
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24 Cont 24 GA+alpha 24 GA+beta
EPB1
Chitinase class I
HvBWMK1
Hypothetical protein
GAPDH
polyA-binding protein
B
HvWRKY51 HvEsi47C
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lativ
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vel
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)Cont -GalY -GlcY
+GA3
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rans
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ontr
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Cont -GalY -GlcY
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evel
(t
rans
crip
t of c
ontr
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100
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Cont -GalY -GlcY
+GA3
Cont -GalY -GlcY
+GA3
Cont -GalY -GlcY
+GA3
Cont -GalY -GlcY
+GA3
Cont -GalY -GlcY
+GA3
Cont -GalY -GlcY
+GA3
Cont -GalY -GlcY
+GA3
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