Lim et al, Supplemental Figure S Arsenic Plant height (Cm) As[μM] b/c g f e d c/d a/b a c/d a a/b Cadmium [mg/Kg] Cd[μM] Plant height (Cm) c g f e d/e d d c a b b a/b Figure S1. Phenotypic effect of rice plants under As and Cd treatments. Rice (cv. Donganbyeo) seeds were germinated, and then grown in soil in the absence or presence of the indicated concentrations of As(V) (Na 2 HAsO 4 ) or Cd (CdSO 4 ) in a controlled environment with a 16 h light/8 h dark photoperiod at 22°C/18°C. Plant height was adjusted using the measuring rule at 4 weeks after transplanting. Error bars represent the standard error, and the same letters above each bar indicate no difference with treatment at a 0.05 significance level of the Duncan’s test. Data are from 3–4 independent assays.

Lim et al, Supplemental Figure S Hydrophobicity index a.a Figure S2. Hydropathy plot of the predicted OsHIR1 protein sequence. The hydropathy plot (mean values for a window of 17 amino acids) was elaborated using the Kyte– Doolittle (KD) hydrophobicity index (

AtUBC21 OsHIR1 35S:EYFP# 1 # 11 # 12 35S:OsHIR1-EYFP Lim et al, Supplemental Figure S3. Figure S3. Semi-quantitative RT-PCR of OsHIR1-overexpressing Arabidopsis and control plants. Total RNAs from 2-week-old seedlings of 35S:OsHIR1-EYFP and 35S:EYFP (empty vector) T 3 transgenic plants were used for RT-PCR. AtUBC21 was used for the internal control.

CloneQDO/X/AX-α-Gal activityTIGR annotation (Rice Genome Annotation Resource)Predicted subcellular localizationTIGR Locus ++++sucrose-phosphate synthase Cytosol LOC_Os01g jacalin-like lectin domain containing protein Cytosol LOC_Os01g universal stress protein domain containing protein Chloroplast LOC_Os05g tRNA synthetases class II domain containing protein Nuclear LOC_Os02g elongation factor Tu Cytosol LOC_Os03g ADP-ribosylation factor Cytosol LOC_Os03g ferredoxin 1-like isoform 1 Mitochodria LOC_Os09g expressed protein Chloroplast LOC_Os09g aquaporin protein Cytosol LOC_Os05g DnaK family protein Cytosol LOC_Os11g mitochondrial-processing peptidase subunit mitochondrial precursor Mitochondria LOC_Os03g Ubiquitin family domain containing protein Cytosol LOC_Os03g NADH dehydrogenase flavoprotein 2 mitochondrial precursor Chloroplast LOC_Os05g OTU-like cysteine protease family protein Nuclear LOC_Os02g VTC2 Chloroplast LOC_Os12g phosphosulfolactate synthase-related protein Cytosol LOC_Os06g ras-related protein Nuclear LOC_Os01g expressed protein Cytoskeleton LOC_Os03g transaldolase Chloroplast LOC_Os01g trehalose-6-phosphate synthase Nuclear LOC_Os09g expressed protein Nuclear LOC_Os06g WD domain G-beta repeat domain containing protein Mitochondria LOC_Os08g pollen-specific protein SF21 Chloroplast LOC_Os06g hypersensitive-induced response protein Cytosol LOC_Os08g CHIT3 - Chitinase family protein precursor Nuclear LOC_Os04g ubiquitin family protein Cytosol LOC_Os02g bZIP transcription factor domain containing protein Nuclear LOC_Os07g PRLI-interacting factor A Mitochodria LOC_Os02g calvin cycle protein CP12 Chloroplast LOC_Os03g receptor-like protein kinase HAIKU2 precursor Plasma membrane LOC_Os12g N-carbamoylputrescine amidase Cytosol LOC_Os02g33080 Lim et al, Supplemental Figure S4. Figure S4. Positive interacting substrate proteins of the OsHIR1 RING E3 ligase. A yeast two-hybrid screening and their predicted subcellular distribution are presented. “+” indicates cell growth on QDO/X/A or density of α-GAL activity (“+,” weak; “++,” strong; “+++,” very strong). Subcellular localizations of positive interacting proteins with OsHIR1 were predicted using the WoLF PSORT program (

Lim et al, Supplemental Figure S5. 35S::OsTIP4;1-EGFP 35S::OsHIR1-EGFP /35S::OsTIP4;1-EGFP 35S::EGFP OsHIR1 OsTIP4;1 Nt18S rRNA AB Figure S5. Coexpression of OsHIR1 and OsTIP4;1 in N. benthaminana leaves. A, Semi-quantitative RT-PCR was conducted to evaluate expression level of OsTIP4;1 in the present or present of OsHIR1 or. Each Agrobacterium (GV3101) containing the35S:OsHIR1-EGFP and 35S:OsTIP4:1-EGFP was transiently co-expressed in N. benthamiana leaves. The 35S:EGFP construct was used as negative control and the Nt18S rRNA gene was used as a internal control. B, The volume integration value of 35S:OsTIP4;1. Autoradiographs were digitized using a Bio-Rad Gel Doc XR+ system. The volume integration value for each band was determined after subtraction of local background by using Molecular Analyst Software (Bio-Rad) version 5.0. Bar indicates standard error of three replicate experiments.