Supplemental Figure 1. Phylogenetic tree of eukaryotic MIPS proteins.
Supplemental Data. Luo et al. (2011). Plant Cell 10.1105/tpc.111.083337
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Supplemental Figure 2. Expression patterns of the three MIPS genes in different
Arabidopsis tissues.
More than 20 transgenic lines were analyzed for each MIPS promoter. Bars = 1 mm.
Supplemental Data. Luo et al. (2011). Plant Cell 10.1105/tpc.111.083337
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Supplemental Figure 3. Phenotypes of mips double and triple mutants and their
Trypan blue staining assays.
(A) to (E) Phenotype of seedlings grown in 16 hr light/8 hr dark growth condition.
Bars = 5 mm.
(F) to (H) Trypan blue staining of wild type (F), mips1 mips2+/- (G), and mips1 mips3
(H) rosette leaves. Dead cells were stained blue. Bars = 1 mm.
Supplemental Data. Luo et al. (2011). Plant Cell 10.1105/tpc.111.083337
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Supplemental Figure 4. PI#1 localization in globular stage embryos of wild-type
and mips1 mips3 double mutant upon BFA treatment.
PIN1 localization in globular stage embryos for wild-type (B) and mips1 mips3
double mutant (D) after 10 min in 50 µM BFA (Invitrogen, Molecular Probes) treatment.
An equal amount of DMSO in 6% glucose solution was used as a negative control for
wild-type (A) and mips1 mips3 (C). Bars = 10 µm.
Supplemental Data. Luo et al. (2011). Plant Cell 10.1105/tpc.111.083337
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Supplemental Figure 5. #umbers of trafficking vesicles in wild type, mips1 mips3
double mutant and PIS2-OX embryo cells.
Cells (n=15 for each line) with similar area of the cross section were used for
calculation. Error bars indicate standard error of mean.
Supplemental Data. Luo et al. (2011). Plant Cell 10.1105/tpc.111.083337
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Supplemental Table 1. Primers used in this study.
T-D#A insertion
MIPS1 sense AGA ACA TTG TGC CAC TCC CTG GG
antisense CCT GAT CGC CAT ATC AAT TAG ACC
MIPS2 sense CGC TAT GGC TAG AGC TAG AGT TCT
antisense CGA TAT AGC CAA TTC GAT AAG ACC
MIPS3 sense GGA GGA TGG GAC ATA AGT GAC ATG
antisense TGA CAT GAG ATT CTC CGT CGT GTC
LBa1 TGG TTC ACG TAG TGG GCC ATC G
DL3 GCT TTC GCC TAT AAA TAC GAC GG
Real Time PCR
MIPS1 sense CGT TGG GAT GAA CGA TAC AAT
antisense ACA TTG TTC CTG ATC GCC ATA
MIPS2 sense GAT CAT GGA GTA CAA GTG AAG G
antisense TAA ACC GAC CAG ATC CAT ATC
MIPS3 sense ATG ATT CTC GAG TAC AAG TGA TC
antisense CGC GAG ATA CCAT TAC ATA TT
TUB2 sense GTT CTC GAT GTT GTT CGT AAG
antisense TGT AAG GCT CAA CCA CAG TAT
RT PCR
MIPS1 sense CGA CTT GCA GAA ACA GCT CAG G
antisense CGC AAC ATA CGG TAC ATA CTT
MIPS2 sense TCC TTG TTG GAG CTG GCA TT
antisense AAA CTT CCC CTC TCC TTC AGC
MIPS3 sense GCT GGT GTT ATC GCC AAT CGT
antisense CCT CAT CCT TGT CCA AAG ATG A
TUB2 sense GTT CTC GAT GTT GTT CGT AAG
antisense TGT AAG GCT CAA CCA CAG TAT
Promoter clone
MIPS1 sense CCA GCA CAT CAA CAT CAC CCT
antisense CTC AAT AAA CAT TTT TGC AGA G
MIPS2 sense TGT CGA TGC GGG ACG TTG AAC
antisense CTC GAT GAA CAT TTT CTC TGG
MIPS3 sense GGGG ACA AGT TTG TAC AAA AAA GCA GGC
TGA TGA AAT GTC TTT GTC CTG GT
antisense GGGG AC CAC TTT GTA CAA GAA AGC TGG
GTA GAA TTA ATT GAT GCT TGA GAG
CDS clone
MIPS1 sense GGGG ACA AGT TTG TAC AAA AAA GCA GGC
TAT ATG TTT ATT GAG AGC TTC AA
antisense GGGG AC CAC TTT GTA CAA GAA AGC TGG
Supplemental Data. Luo et al. (2011). Plant Cell 10.1105/tpc.111.083337
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GTT TCA CTT GAA TTC CAT GAT CA
MIPS2 sense GGGG ACA AGT TTG TAC AAA AAA GCA GGC
TAT ATG TTC ATC GAG AGC TTC AA
antisense GGGG AC CAC TTT GTA CAA GAA AGC TGG
GTT TCA CTT GTA CTC CAT GAT CA
MIPS3 sense GGGG ACA AGT TTG TAC AAA AAA GCA GGC
TAT ATG TTC ATC GAA AGC TTC AA
antisense GGGG AC CAC TTT GTA CAA GAA AGC TGG
GTT TCA CTT GTA CTC GAG AAT CA
INO1 sense GGGG ACA AGT TTG TAC AAA AAA GCA GGC
TCA ATG ACA GAA GAT AAT AT
antisense GGGG AC CAC TTT GTA CAA GAA AGC TGG
GTA TTA CAA CAA TCT CTC TTC
Over expression
PIS1 sense GGGG ACA AGT TTG TAC AAA AAA GCA GGC
TAA AGA GAT TCT CAC GAT CG
antisense GGGG AC CAC TTT GTA CAA GAA AGC TGG
GTT TTG GTA ACG AGA TAA TGC TGA
PIS2 sense GGGG ACA AGT TTG TAC AAA AAA GCA GGC
TTT CCT GAC AGC GAG AAG AA
antisense GGGG AC CAC TTT GTA CAA GAA AGC TGG
GTA ACA CGA TTC ACG GGC ACA
UGT84B1 sense GGGG ACA AGT TTG TAC AAA AAA GCA GGC
TAG CCT TGA AGA AGA AGA AA
antisense GGGG AC CAC TTT GTA CAA GAA AGC TGG
GTC CTT GAT AAT GGT CAG CTA
OX lines T-D#A
insertion
PIS1 sense GTG TTT GGA ATC GGA GTA AGG A
PIS2 sense TTT TCT CCT GGC TTT GAC ATT G
UGT84B1 sense CTC ATA GCA ACG AAC CAA ACC
vector antisense GTT CAC CTT GAT GCC GTT CTT
Annotation:
MIPS1 sense/LBa1, MIPS2 antisense/LBa1, MIPS3 sense/DL3 primer pairs were used for mips
T-DNA insertion detection.
For the construction of gateway over expression vector, pH7FWG2 was digested by HindIII/SpeI
to release 35S promoter and filling in with the tetramer-35S-enhancer fragment.
Supplemental Data. Luo et al. (2011). Plant Cell 10.1105/tpc.111.083337
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Supplemental Table 2. Accession numbers for the sequences used in the
alignment in Supplemental Figure 1 and Supplemental Dataset 1.
Species Abbreviation GI
Saccharomyces cerevisiae SceINO1 37362665
Ostreococcus tauri Ota 116061703
Arabidopsis thaliana (At5g10170) AthMIPS3 15236025
Brassica rapa subsp. Pekinensis Bra 72384416
Triticum aestivum Tae 14548098
Nicotiana tabacum Nta 14548096
Nicotiana paniculata Npa 14548099
Sesamum indicum Sin 14548095
Arabidopsis thaliana (At2g22240) AthMIPS2 15227169
Brassica napus MIPS1 BnaMIPS1 14548093
Brassica napus MIPS2 BnaMIPS2 215274557
Brassica napus MIPS3 BnaMIPS3 215274559
Brassica napus MIPS4 BnaMIPS4 215274561
Brassica napus MIPS5 BnaMIPS5 215274563
Brassica napus MIPS6 BnaMIPS6 215274565
Glycine max (1) GmaMIPS1 119960405
Glycine max (2) GmaMIPS2 84311235
Glycine max (3) GmaMIPS3 84311237
Glycine max (4) GmaMIPS4 84311239
Passiflora edulis f. flavicarpa Ped 95104695
Arabidopsis thaliana (AT4g39800) AthMIPS1 15236025
Suaeda maritima subsp. Salsa Sma 16755889
Medicago truncatula Mtr 92898152
Phaseolus vulgaris (1) Pvu_1 14548092
Phaseolus vulgaris (2) Pvu_2 60417544
Avicennia marina Ama 13447457
Spirodela polyrrhiza Spo 1170568
Zea mays (1) Zma_1 11762100
Zea mays (2) Zma_2 14548094
Supplemental Data. Luo et al. (2011). Plant Cell 10.1105/tpc.111.083337
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Trifolium pratense Tpr 84468396
Avena sativa Asa 18181983
Xerophyta viscosa Xvi 32492882
Oryza sativa (japonica) (Os03g0192700) Osa_1 115451283
Oryza sativa (japonica) (Os10g0369900) Osa_2 115481722
Hordeum vulgare Hvu 14548089
Citrus x paradisi Cpa 1170567
Physcomitrella patens (1) Ppa_1 168055751
Physcomitrella patens (2) Ppa_2 168020111
Phaeodactylum tricornutum Ptric 219116747
Brassica juncea Bju 165929042
Picea sitchensis Psi 148907091
Vitis vinifera Vvi 157345093
Cicer arietinum Car_1 198401802
Cicer arietinum Car_2 198401804
Mesembryanthemum crystallinum Mcr 14548091
Arabidopsis arenosa Aare 217426800
Medicago sativa Msa 139478073
Vigna radiata Vra 189169790
Chlamydomonas reinhardtii Cre 159490574
Thalassiosira pseudonana Tps 220971094
Arthrocnemum glaucum Agl 145315805
Kluyveromyces lactis Kla 50307609
Candida glabrata Cgl 50290163
Debaryomyces hansenii Dha 50426419
Yarrowia lipolytica Yli 50545902
Aspergillus fumigatus Afu 70988765
Neosartorya fischeri Nfi 119497811
Laccaria bicolor Lbi_1 170100589
Laccaria bicolor Lbi_2 170100971
Pyrenophora tritici-repentis Ptrit 189198886
Cryptococcus neoformans Cne 58265488
Supplemental Data. Luo et al. (2011). Plant Cell 10.1105/tpc.111.083337
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Penicillium marneffei Pma 212527356
Sclerotinia sclerotiorum Ssc 156066001
Gibberella zeae Gze 46136849
Drosophila melanogaster Dme 17137626
Aedes aegypti Aae 157132320
Anopheles gambiae Aga 158300873
Paramecium tetraurelia Pte 145499166
Trichomonas vaginalis Tva 123433216
Caenorhabditis briggsae Cbr 157750944
Ustilago maydis Uma 71022933
Malassezia globosa Mgl 164657384
Caenorhabditis elegans Cel 25147617
Tetrahymena thermophila Tth 118364145
Trypanosoma brucei Tbr 71747782
Strongylocentrotus purpuratus Spu 115640689
Coprinopsis cinerea Cci 169846152
Hydra magnipapillata Hma 221129143
Nematostella vectensis Nve 156381404
Mus musculus Mmus 12963757
Bos taurus Bta 114051253
Rattus norvegicus Rno 189163481
Canis familiaris Cfa 73986122
Salmo salar Ssa 213511546
Branchiostoma floridae Bfl 219472225
Macaca mulatta Mmul 109123938
Equus caballus Eca 194232542
Xenopus laevis Xla_1 148233209
Xenopus laevis Xla_2 147901823
Nasonia vitripennis Nvi 156555239
Xenopus (Silurana) tropicalis Xtr 187607836
Apis mellifera Ame 66546786
Dictyostelium discoideum Ddi 66808743
Supplemental Data. Luo et al. (2011). Plant Cell 10.1105/tpc.111.083337
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Macaca fascicularis Mfa 75076346
Homo sapiens HsaMIPS1 7705558
Tribolium castaneum Tca 91088055
Ixodes scapularis Isc 215495900
Pediculus humanus corporis Phu 212515411
Entamoeba histolytica Ehi 67477740
Tetraodon nigroviridis Tni 47229914
Pan troglodytes Ptrog 114676130
Supplemental Data. Luo et al. (2011). Plant Cell 10.1105/tpc.111.083337
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