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Figure S1. Characterization of homozygous T-DNAinsertion lines. (a) Diagram of the T-DNA insertion into theAtpht4;6 gene. For T-DNA screening, the insertion mutantSAIL 809_B01 obtained from the Nottingham Arabidopsis Stock Centre(NASC) was used. Arrows represent binding sites of primers used formolecular characterization. (b) Identification of homozygouspht4;6 plants via PCR. Using primer combinationAt5g44370_11/At5g44370_12 in lines 1, 3 and 5 leads to thepht4;6 product in the Wt and heterozygous line. For identification of T-DNA insertion, the primer combination LB-3/At5g44370_12 was used in lines 2, 4 and 6. (c) Detection of cDNA in homozygous and Wt plants. cDNA of the size of 250 bp was amplified with the primer combination At5g44370_29/At5g44370_30.

Figure S2. Complementation of the pht4;6mutation with the Oryza sativa homologue Os11g08370. (a)Diagram of the construct used for complementation of pht4;6plants. cDNA of the Os11g08370 gene was fused with a HIS-Tag andtransferred in a plant transformation vector under the control ofthe 35-S-promotor. Binding sites of the primers are shown asarrows. (b) Phenotypes of 5-week old Wt, pht4;6 andpht4;6::Os11g08370 plants. (c) Identification ofcomplemented mutant plants via PCR. Using primer combinationAt5g44370_ 11/At5g44370_12 in line 1, 4 and 7 leads to thepht4;6 product in the Wt. For identification of T-DNA insertion, the primer combination LB-3/At5g44370_12 was used in line 2, 5 and 8. The complementation was verified with the primers Os11g08370_4/Os11g08370_5, showing a PCR-product in line 9.

Figure S3. Complementation of the pht4;6mutation with Arabidopsis thaliana At5g44370. (a) Diagram ofthe construct used for complementation of pht4;6 plants.Genomic DNA of the At5g44370 gene including 1257 bp of theendogenous promotor sequence and 361 bp of the 3’UTR regionwas transferred in a plant transformation vector (pGreen0029).Binding sites of primers are shown as arrows. (b) Phenotypes of8-week old Wt, pht4;6 and pht4;6::At5g44370 plants.(c) Identification of complemented mutant plants via PCR. Usingprimer combination At5g44370_11/At5g44370_12 in lines 1, 4 and 7leads to the pht4;6 pro-duct in the Wt. For identification of T-DNA insertion, the primer combination LB-3/At5g44370_12 was used in lines 2, 5 and 8. The complementation was verified with the kanamycin primers nptII_s/nptII_as, showing a PCR-product of 600 bp in line 9.

Figure S4. The PHT4;6 protein locates to thetrans-Golgi. (a, b). Arabidopsis thaliana protoplasts,generated by cell suspension culture (Brightfield), expressing thePHT4;6::GFP fusion protein (a) and Golgi-mannosidase::RFP fusionprotein (b). (c) Merge of both channels with a magnificationof the colocalization of both fusion proteins. (d, e).Arabidopsis thaliana protoplasts, generated by cellsuspension culture, expressing the PHT4;6::GFP fusion protein (d)and the DsRed-tagged sialin transferase (ST) (e). (f) Mergeindicates a colocalization in the trans-Golgi compartment. Analysisof fluorescence proteins was performed by confocal laser scanningmicroscopy. Scale bar = 2 μm.

Table S1. Primer used for identification ofpht4;6 T-DNA insertion lines, com-plementation of pht4;6 with the rice homologue Os11g08370 and for qRT-PCR studies. Restriction sites are displayed in bolt.

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