Supplemental Data. Seo et al. (2014). Plant Cell 10.1105 ......Feb 07, 2014  · Seo et al. (2014)....

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Supplemental Data. Seo et al. (2014). Plant Cell 10.1105/tpc.113.119974 Supplemental Figure 1. Protein alignment of ABD1 from other model organisms. The alignment was performed with H. sapiens DCAF8, M. musculus DCAF8 and O. sativa Os10g0544500. The WD40 domains are underlined. The red triangles indicate the WDxR motif within the WD40 domain and “x” stands for an undefined amino acid. The shading mode indicates the level of conservation, with red letters in black shading corresponding to a high level of conservation (100%), blue letters in dark gray shading corresponding to a moderate level of conservation (80%) and green letters in light gray shading corresponding to a low level of conservation (60%).

Transcript of Supplemental Data. Seo et al. (2014). Plant Cell 10.1105 ......Feb 07, 2014  · Seo et al. (2014)....

Page 1: Supplemental Data. Seo et al. (2014). Plant Cell 10.1105 ......Feb 07, 2014  · Seo et al. (2014). Plant Cell 10.1105/tpc.11.1199743 Supplemental Figure 1. Protein alignment of ABD1

Supplemental Data. Seo et al. (2014). Plant Cell 10.1105/tpc.113.119974

Supplemental Figure 1. Protein alignment of ABD1 from other model organisms. The alignment was performed with H. sapiens DCAF8, M. musculus DCAF8 and O. sativa Os10g0544500. The WD40 domains are underlined. The red triangles indicate the WDxR motif within the WD40 domain and “x” stands for an undefined amino acid. The shading mode indicates the level of conservation, with red letters in black shading corresponding to a high level of conservation (100%), blue letters in dark gray shading corresponding to a moderate level of conservation (80%) and green letters in light gray shading corresponding to a low level of conservation (60%).

Page 2: Supplemental Data. Seo et al. (2014). Plant Cell 10.1105 ......Feb 07, 2014  · Seo et al. (2014). Plant Cell 10.1105/tpc.11.1199743 Supplemental Figure 1. Protein alignment of ABD1

Supplemental Data. Seo et al. (2014). Plant Cell 10.1105/tpc.113.119974

Supplemental Figure 2. abd1-1 and abd1-2 seedlings have no change in expression of RD20 and RD22 after ABA and NaCl treatment. 7-day-old Col-0 wt, abd1-1 and abd1-2 seeds were grown in the absence or presence of 0.5 µM ABA or 100 mM NaCl. mRNA levels were determined by quantitative real-time PCR analysis. Relative amounts of transcripts were normalized to the levels of ACTIN2 within the same sample. (A) RD22 after ABA treatment. (B) RD22 after NaCl treatment. (C) RD20 after ABA treatment. (D) RD20 after NaCl treatment. (E) ABI3 after ABA treatment. Results are from three biological replicates and values represent mean ± SD (n=9). N.S., not significant.

Page 3: Supplemental Data. Seo et al. (2014). Plant Cell 10.1105 ......Feb 07, 2014  · Seo et al. (2014). Plant Cell 10.1105/tpc.11.1199743 Supplemental Figure 1. Protein alignment of ABD1

Supplemental Data. Seo et al. (2014). Plant Cell 10.1105/tpc.113.119974

Supplemental Figure 3. ABD1 does not directly interact with DWA proteins, ABI5 homologs or ABI3 by yeast two hybrid assays. Assays were performed with ABD1 protein as prey and various ABA responsive proteins as bait to monitor their interactions. Empty vector was used as a negative control. Yeast were grown in the presence of X-gal for 15 hours, after which images were taken. (A) Interaction between ABD1 and DWA1, DWA2, or DWA3. (B) Interaction between ABD1 and the five most closely related ABI5 homologs: ABF1, DPBF2, AREB1, AREB3 and AREB1. (C) Interaction between ABD1 and ABI3. (D) Immunoblots of the target fusion proteins in the yeast two hybrid assays in (B) and (C). The LexA fusions were detected by anti-LexA antibodies and the expression of these proteins are indicated by the arrows. Empty LexA vector was used as a negative control. Ponceau S staining was used as a loading control. A total of 15 µg of protein was loaded in each lane.

Page 4: Supplemental Data. Seo et al. (2014). Plant Cell 10.1105 ......Feb 07, 2014  · Seo et al. (2014). Plant Cell 10.1105/tpc.11.1199743 Supplemental Figure 1. Protein alignment of ABD1

Supplemental Data. Seo et al. (2014). Plant Cell 10.1105/tpc.113.119974

Supplemental Figure 4. Quantification of ABI5 protein degradation after ABA removal. Immunoblot assays shown in Figure 9 were quantified showing relative amounts of ABI5 protein in Col-0 wt and abd1-1 seeds that were treated with 5 µM ABA in white light for 3 days, and then harvested at the indicated times after the removal of ABA with either being washed out with (A) liquid media, (B) liquid media supplemented with the proteasome inhibitor MG132 (50 µM) or (C) the protein synthesis inhibitor CHX (100 µM). Relative amounts of ABI5 protein were normalized to the levels of RPN6 within the same sample. Values are means ± SD (n=3). Significant difference was determined by a Student’s t-test; single or double stars indicate a P values of P<0.03 or P<0.004, respectively.

Page 5: Supplemental Data. Seo et al. (2014). Plant Cell 10.1105 ......Feb 07, 2014  · Seo et al. (2014). Plant Cell 10.1105/tpc.11.1199743 Supplemental Figure 1. Protein alignment of ABD1

Supplemental Data. Seo et al. (2014). Plant Cell 10.1105/tpc.113.119974

Supplemental Tables Supplemental Table 1. List of primers used for genotyping and RT-PCR analysis. Name T-DNA Line Forward Primer (5'-3') Reverse Primer (5'-3') Genotypinga RT-PCR

ABD1 (At4g38480)

SALK_051074 (abd1-1)

GAAAAGGCCGAGGACCGACCAT CTGTCATCAGAACCAGAAAG FP1+RP1

(F/LBb1.3) FP1+RP1

ABD1 (At4g38480)

SAIL_648_G02 (abd1-2)

CTTTCTGGTTCTGATGACAGA ATGTCTGTCTGCTTCCATGG FP2+RP2 (F/LB3) FP2+RP2

GAAAAGGCCGAGGACCGACCAT TATTATTGAAGTGGCCGGAA FP1+RP3

LBb1.3 ATTTTGCCGATTTCGGAAC

LB3 TAGCATCTGAATTTCATAACCAATCTCGATACAC

RPN6 AAGGCACGATAGATCTGCAGA TCGAGGGTAGCCGAGTAGAT

a primer sets for confirmation of homo- or heterozygous lines

Page 6: Supplemental Data. Seo et al. (2014). Plant Cell 10.1105 ......Feb 07, 2014  · Seo et al. (2014). Plant Cell 10.1105/tpc.11.1199743 Supplemental Figure 1. Protein alignment of ABD1

Supplemental Data. Seo et al. (2014). Plant Cell 10.1105/tpc.113.119974

Supplemental Table 2. List of primers used in quantitative real-time PCR analysis. Name Forward Primer (5'-3') Reverse Primer (5'-3')

ABD1 TGGGTCTCTTACCCAATCGCAGTT CAGAAGCAGTCTGCCAATCCCAAA

RD29A GGAGGAGAAGAAGAGAAGAA TAAAGCTCCTTCTGCACCGG

RD29B ACGGAAACATCGGACTGG TCCGTTGACCACCGAGAT

RD22 TTCGGAAGAAGCGGAGAT CAGTGGAAACAGCCCTGA

RD20 CGTTTCAAACAAAGTTGAATGGATAC

TTAGTGCTTGTTTGCGAGAATTGGCC

ABI5 AACATGCATTGGCGGAGT TTGTGCCCTTGACTTCAAACT

ABI3 TCTTGAATGGGTCCAAAC AGGGTTTTGAAATGGATC

ACTIN2 CAAGGCCGAGTATGATGAGG GAAACGCAGACGTAAGTAAAAAC

Page 7: Supplemental Data. Seo et al. (2014). Plant Cell 10.1105 ......Feb 07, 2014  · Seo et al. (2014). Plant Cell 10.1105/tpc.11.1199743 Supplemental Figure 1. Protein alignment of ABD1

Supplemental Data. Seo et al. (2014). Plant Cell 10.1105/tpc.113.119974

Supplemental Table 3. List of primers used for LCI analysis. Name Forward Primer (5'-3') Reverse Primer (5'-3') ABD1_KpnI_cLUC

GGGGTACCATGAGTGGAAGGCCGGCG

GGGGTACCTCATCCCTCATCCCCATCCCC

ABI5_KpnI_nLUC

GGGGTACCATGGTAACTAGAGAAACGAAGTTG

GGGGTACCGAGTGGACAACTCGGGTTC

Page 8: Supplemental Data. Seo et al. (2014). Plant Cell 10.1105 ......Feb 07, 2014  · Seo et al. (2014). Plant Cell 10.1105/tpc.11.1199743 Supplemental Figure 1. Protein alignment of ABD1

Supplemental Data. Seo et al. (2014). Plant Cell 10.1105/tpc.113.119974

Supplemental Table 4. List of primers used for yeast two-hybrid analysis. Name Sequence (5'-3')

ABD1_EcoRI_Forward GGGAATTCTCATCCCTCATCCCCATC

ABD1_EcoRI_Reverse CGGAATTCATGAGGGAAGGCCGGC

AREB1_EcoRI_Forward CCGGAATTCATGGATGGTAGTATGAATTTGGGG

AREB1_NotI_Reverse AAGGAAAAAAGCGGCCGCTCACCAAGGTCCCGACTC

AREB2_EcoRI_Forward CCGGAATTCATGGGAACTCACATCAATTTCAAC

AREB2_NotI_Reverse AAGGAAAAAAGCGGCCGCTCACCATGGTCCGGTTAATGTCC

DWA1_EcoRI_Forward CGGAATTCATGTACGGAGACGCTACAAAC

DWA1_EcoRI_Reverse CGGAATTCTTACAATGAACTGCTACGAA

DWA2_XhoI_Forward CCGCTCGAGATGCAAGGAGGATCATCGGG

DWA2_XhoI_Reverse CCGCTCGAGCTATCTTCTTGGCAGGAAAG

DWA3_EcoRI_Forward CGGAATTCATGGCGAAGCGTGGTTATAA

DWA3_EcoRI_Reverse CGGAATTCCTATGGTGTTTGAAGAACTA

ABI5_EcoRI_Forward CGGAATTCGGATGGTAACTAGAGAAACGAAG

ABI5_EcoRI_Reverse CGGAATTCTTAGAGTGGACAACTCGGGT

ABF1_BamHI_Forward CGGGATCCCGATGGGTACTCACATTGATATCAAC

ABF1_NotI_Reverse AAGGAAAAAAGCGGCCGCTCACCTTCTTACCACGGACC

DPBF2_BamHI_Forward CGGGATCCCGATGTCGGTTTTCGAATCGGAGACTTCG

DPBF2_NotI_Reverse AAGGAAAAAAGCGGCCGCTTACCACCCGGCACTGGCCA

AREB3_BamHI_Forward CGGGATCCCGATGGATTCTCAGAGGGGTATTG

AREB3_NotI_Reverse AAGGAAAAAAGCGGCCGCTCAGAAAGGAGCCGAGCTTG

ABI3_BamHI_Forward CGGGATCCCGATGAAAAGCTTGCATGTGGCG

ABI3_NotI_Reverse AAGGAAAAAAGCGGCCGCTCATTTAACAGTTTGAGAAGTTGG

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Supplemental Data. Seo et al. (2014). Plant Cell 10.1105/tpc.113.119974

Supplemental Table 5. List of primers used for BiFC assay. Name Sequence (5'-3')

ABD1_NotI_ pSY728_Forward TTGCGGCCGCAAATGAGTGGAAGGCCGGCGAAA

ABD1_NotI_ pSY728_Reverse TTGCGGCCGCTTTCCCTCATCCCCATCCCCAT

ABI5_BamHI_ pSY735_Forward CGGGATCCCGATGGTAACTAGAGAAACGAAGTTG

ABI5_BamHI_ pSY735_Reverse CGGGATCCTTAGAGTGGACAACTCGGGTTC

DDB1a-NotI_ pSY738_Forward TTGCGGCCGCAAATGAGCTCATGGAACTACGTTG

DDB1a-NotI_ pSY738_Reverse TTGCGGCCGCTTGTGAAGCCTAGTGAGTTCTTCAA


Supplemental Data. De Clercq et al. Plant Cell. (2013). … Data. De Clercq et al. Plant Cell. (2013). 10.1105/tpc.113.117168 1 Supplemental Figure 1. Binding of NAC transcription

Supplemental Data. De Clercq et al. Plant Cell. (2013). … Data. De Clercq et al. Plant Cell. (2013). 10.1105/tpc.113.117168 1 Supplemental Figure 1. Binding of NAC transcription

E Wild -type of 35S:MYB106-SRDX F flowers and torenia leaf. · PDF fileSupplemental Data. Oshima et al. (2013). Plant Cell 10.1105/tpc.113.110783 1 / 18 Supplemental Figure 1. Cuticle

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Supplemental Data. Xia et al. Plant Cell. (2013). …...2013/05/09  · Supplemental Data. Xia et al. Plant Cell. (2013). 10.1105/tpc.113.110957 2& & & Supplemental Figure 2. The miRNA-TASL-PPR-siRNA

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Supplemental Data. Adachi et al. (2015). Plant Cell …...2015/09/02  · Supplemental Data. Adachi et al. (2015). Plant Cell 10.1105/tpc.15.00213 Supplemental Figure 1. Promoter Activity

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Supplemental Data. Hsu et al. Plant Cell. (2013). 10.1105 ...€¦ · Supplemental Data. Hsu et al. Plant Cell. (2013). 10.1105/tpc.113.114447 1 . Supplemental Figure 1. Induction

Supplemental Data. Hsu et al. Plant Cell. (2013). 10.1105 ...€¦ · Supplemental Data. Hsu et al. Plant Cell. (2013). 10.1105/tpc.113.114447 1 . Supplemental Figure 1. Induction

Supplemental Data. Lei$etal.$(2013).$PlantCell$10.1105/tpc ... · 12/23/2013  · Supplemental Data. Lei$etal.$(2013).$PlantCell$10.1105/tpc.113.116715$ 4 Supplemental Figure 5. Similar

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SUPPLEMENTAL FIGURES - The Plant Cell · 3/14/2014  · Supplemental Data. Morreel et al. Plant Cell (2014). 10.1105/tpc.113.122242. 1 SUPPLEMENTAL FIGURES Supplemental Figure 1.

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Supplemental Data. Geng et al. (2013). Plant Cell 10.1105 ... · Supplemental Data. Geng et al. (2013). Plant Cell 10.1105/tpc.113.112896 9" " Supplemental Figure 9. The ProRAB18:GFP

Supplemental Data. Geng et al. (2013). Plant Cell 10.1105 ... · Supplemental Data. Geng et al. (2013). Plant Cell 10.1105/tpc.113.112896 9" " Supplemental Figure 9. The ProRAB18:GFP

Supplemental Data. Gong et al. (2012). Plant Cell 10.1105 ... · 9/11/2012  · Plant Cell 10.1105/tpc.111.100511 . Supplemental Figure 2. Fine structure of the functional core of

Supplemental Data. Gong et al. (2012). Plant Cell 10.1105 ... · 9/11/2012  · Plant Cell 10.1105/tpc.111.100511 . Supplemental Figure 2. Fine structure of the functional core of

20121027 Supplemental Data Final - Plant Cell · Supplemental Data. Ren et al. (2012). Plant Cell 10.1105/tpc.111.107144 2 Supplemental figure 1. Functional analysis of BP12 lines

20121027 Supplemental Data Final - Plant Cell · Supplemental Data. Ren et al. (2012). Plant Cell 10.1105/tpc.111.107144 2 Supplemental figure 1. Functional analysis of BP12 lines

Supplemental Figure 1. ABA responses of SWI2/SNF2 chromatin remodeling mutants… · 2012-12-03 · Supplemental Data. Han et al. Plant Cell. (2012). 10.1105/tpc.112.105114 1 Supplemental

Supplemental Figure 1. ABA responses of SWI2/SNF2 chromatin remodeling mutants… · 2012-12-03 · Supplemental Data. Han et al. Plant Cell. (2012). 10.1105/tpc.112.105114 1 Supplemental

Supplemental Data. Cubillos et al. (2014). Plant Cell 10 ... · Supplemental Data. Cubillos et al. (2014). Plant Cell 10.1105/tpc.114.130310 Supplemental Figure 4. Overlap comparison

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Supplemental Data. Seo et al. (2017). Plant Cell 10.1105 ...€¦ · Supplemental Data. Seo et al. (2017). Plant Cell 10.1105/tpc.16.00886 A B ORF No. Size (bp) aa No. 1185 2 132

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Supplemental Data. Antoniadi et al. (2015). Plant Cell. 10 ......Jul 10, 2015  · Antoniadi et al. (2015). Plant Cell. 10.1105/tpc.15.00176 Supplemental Figure 5. Cytokinin Metabolism

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Data. Mbengueet al. (2010). Plant Cell 10.1105/tpc.110.075861...Supplemental Figure 6. PUB1-RNAi Construct Efficiency. The PUB1-RNAi construct efficiently down-regulates PUB1 gene

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