AtTIL-P91V. AtTIL-P92V. AtTIL-P95V. AtTIL-P98V YFP-HPR

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Online Resource 1. Primers used to generate constructs AtTIL-P91V, AtTIL-P92V, AtTIL-P95V and AtTIL-P98V and YFP(HPR) using overlapping PCR. pentr/d- TOPO-AtTIL was used as template to generate the constructs AtTIL-P91V, AtTIL- P92V, AtTIL-P95V and AtTIL-P98V. pentr/d-topo-yfp was used as template to generate YFP-HPR construct. Construct AtTIL-P91V AtTIL-P92V AtTIL-P95V AtTIL-P98V YFP-HPR Primer AtTIL(P91V)B: GGA GGA AAG GAA CGA CAT AGA AC AtTIL(P91V)C: GTT CTA TGT CGT TCC TTT CCT CC AtTIL(P92V)B: GAT TGG GAG GAA AAC AGG GAC ATA G AtTIL(P92V)C: CTA TGT CCC TGT TTT CCT CCC AAT C AtTIL(P95V)B: CGG GAA TGA TTA CGA GGA AAG GAG AtTIL(P95V)C: CTC CTT TCC TCG TAA TCA TTC CCG AtTIL(P98V)B: GTC TCC GGT GAC GAC AAT GAT TGG G AtTIL(P98V)C: CCC AAT CAT TGT CGT CAC CGG AGA C YFP(HPR)B: CAG GAA TTA TAG GCA AAA ATG GTG GAA CCT TGT CGG CCA TGA TAT AGA CG YFP(HPR)C: TCC ACC ATT TTT GCC TAT AAT TCC TGT TAA GAA CGG CAT CAA GGT GAA C

Online Resource 2. Sequence alignment of the HPR motif of plant TIL proteins. TILs were identified using the AtTIL protein sequence as a query using BLASTP against the GenBank non-redundant protein sequence database. Only full-length sequences have been included. Amino acid residues common to all sequences are shown in black boxes. In the consensus sequence, Φ corresponds to hydrophobic residues and X to any amino acid residue. Name Species Accession No. HPR motif sequence AtTIL Arabidopsis thaliana gi-15242942 YVPPFLPIIPVT BdTIL1 Bracunkodium distachyon gi-357147915 YVPPFLPVFPVV BdTIL2 Bracunkodium distachyon gi-357150111 YVPPILPIIPVV BnTIL Brassica napus gi-77744889 YVPPFLPIIPVT BrTIL Brassica rapa gi-157849740 YVPPFLPIIPVT CaTIL Capsicum annuum gi-50236424 YVPPFLPVIPVV CsTIL Citrus sinensis gi-77744899 YVPPFFPIIPVV EgTIL Elaeis guineensis gi-192911934 YVPPFFPIIPVT GaTIL Gossypium arboreum gi-77744897 YVPPFLPIIPIV GaTIL2 Gossypium arboreum gi-77744869 YVPPFLPIIPVT GmTIL1 Glycine max gi-351734470 WVPPFLPIIPVT GmTIL2 Glycine max gi-351724275 YIPPFLPIIPIN GmTIL3 Glycine max gi-351726387 WVPPFLPLFPVT GmTIL4 Glycine max gi-351721591 YVPPFLPIIPVT HvTIL1 Hordeum vulgare gi-77744845 YVPPFLPIIPVV HvTIL2 Hordeum vulgare gi-326527227 YVPPFLPVFPVT HvTIL3 Hordeum vulgare gi-77744853 YVPPFLPVFPVT LjTIL1 Lotus japonicus gi-388518293 YVPPFLPIIPVV LjTIL2 Lotus japonicus gi-388522945 FVPPFLPIIPVV McTIL1 Mesembryanthemum crystallinum gi-77744895 YVPPFLPIIPVT McTIL2 Mesembryanthemum crystallinum gi-77744867 YVPPFLPIIPVT MtTIL1 Medicago truncatula gi-357480181 LVPPFLPFIPAV MtTIL2 Medicago truncatula gi-357480173 YVPPFLPIIPAV MtTIL3 Medicago truncatula gi-357480171 YVPPMLPIIPVT OsTIL1 Oryza sativa japonica gi-115447273 YLPPFLPVIPVV OsTIL2 Oryza sativa japonica gi-115476610 YVPPFLPIFPVV OsTIL3 Oryza sativa japonica gi-151935409 YLPPFLPVIPVV PaTIL Prunuspersica gi-77744891 YVPPFLPIIPVV PbTIL Populus balsamifera gi-77744901 YVPPFLPIIPVV PeTIL Populus euphratica gi-209967467 YVPPFLPIIPVV PpTIL Physcomitrella patens gi-168066921 LVPPFFPIFPVT PsTIL Piceasitchensis gi-116783015 MVPPFLPIIPVY PtreTIL Populus tremuloides gi-77744903 YVPPFLPIIPVV PtriTIL Populus trichocarpa gi-224143988 YVPPFLPIIPVV PtTIL Pinus taeda gi-77744881 MVPPFFPIIPVY RcTIL Ricinus communis gi-255565025 YVPPFLPIIPVV SbTIL1 Sorghum bicolor gi-242065756 YLPPFLPVIPVV SbTIL2 Sorghum bicolor gi-77744857 YVPPFLPVFPVT SbTIL3 Sorghum bicolor gi-242079289 YVPPFLPVFPVT SlTIL1 Solanum lycopersicum gi-350539735 YVPPFLPIIPVT SlTIL2 Solanum lycopersicum gi-350539918 YIPPFLPIIPIV SmTIL1 Selaginella moellendorffii gi-302802027 WVPPFLPVFPVT SmTIL2 Selaginella moellendorffii gi-302818492 WVPPFLPVFPVT SoTIL Saccharum officinarum gi-77744875 YLPPFLPIIPVV StTIL1 Solanum tuberosum gi-77744887 YVPPFLPIIPVT StTIL2 Solanum tuberosum gi-77744865 YVPPFLPIIPVT SuTIL Syntrichia ruralis gi-77744919 MVPPFLPVIPVT TaTIL1 Triticuma estivum gi-18650668 YVPPFLPIIPVV TaTIL2 Triticuma estivum gi-77744851 YVPPFLPVFPVT VvTIL Vitis vinifera gi-77744883 YVPPFLPIIPVV ZmTIL1 Zea mays gi-226530914 YLPPFLPIVPVV ZmTIL2 Zea mays gi-226529802 YVPPFLPLIPVT Consensus XΦPPΦΦPΦΦPΦX

Online Resource 3. Hydropathy plot and model of the tertiary structure of SlTIL1 and SlTIL2. a Hydropathy plot (Kyte and Doolittle) of SlTIL1 showing the position and primary sequence of the HPR motif. b Model of the tertiary structure of SlTIL1 obtained using the Swiss-Model Program (Arnold et al., 2006). E. coli BCL protein (PDB ID:2ACO Chain A) was used as template. The dashed line indicates the axis of the lipocalin β-barrel. c Hydropathy plot (Kyte and Doolittle) of SlTIL2 showing the position and primary sequence of the HPR motif. d Model of the tertiary structure of SlTIL2 obtained using the Swiss-Model Program (Arnold et al., 2006). E. coli BCL protein (PDB ID:2ACO Chain A) was used as template. The dashed line indicates the axis of the lipocalin β-barrel.

Online Resource 4. Subcellular localization of SlTIL1 and SlTIL2 in agroinfiltrated N benthamiana leaves. N. benthamiana leaves were agroinfiltrated for the transient expression of: a YFP:SlTIL1, b SlTIL1:YFP, c YFP:SlTIL2 and d SlTIL2:YFP. Cells were imaged at 3 days post infiltration. Images are reconstructed by superposition of series of confocal optical sections. Scale bar = 50 μm.

Online Resource 6. Co-localization of SlTIL1:YFP with cell markers in agroinfiltrated N benthamiana leaves. N. benthamiana leaves were agroinfiltrated for the co-expression of SlTIL:YFP and markers for plasma membrane (PM), tonoplast (TO), endoplasmic reticulum (ER), peroxisome (PO), mitochondria (MIT), Golgi (G) and plastids (Cl). Numbers on the lefthand side correspond to: 1 fluorescence of CFP intracellular markers, 2 fluorescence of SlTIL:YFP, 3 merge of images from 1 and 2, and 4 bright field. Each image corresponds to a single confocal optical section. Scale bar = 10 μm.

Online Resource 7. Co-expression of YFP(HPR) with cell markers in agroinfiltrated N benthamiana leaves. N. benthamiana leaves were agroinfiltrated for the co-expression of YFP-HPR with of SlTIL:YFP and markers for plasma membrane (PM), tonoplast (TO), endoplasmic reticulum (ER), peroxisome (PO), mitochondria (MIT), Golgi (G) and plastids (Cl). Numbers on the left-hand side correspond to: 1 fluorescence of CFP intracellular markers, 2 fluorescence of YFP-HPR, 3 merge of images from 1 and 2, and 4 bright field. Each image corresponds to a single confocal optical section. Scale bar = 10 μm.

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