Supplemental Data. Luo et al. (2011). Plant Cell 10.1105/tpc.111.083337
Supplemental Figure 1. Phylogenetic tree of eukaryotic MIPS proteins.
1
Supplemental Data. Luo et al. (2011). Plant Cell 10.1105/tpc.111.083337
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.
2
Supplemental Data. Luo et al. (2011). Plant Cell 10.1105/tpc.111.083337
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.
3
Supplemental Data. Luo et al. (2011). Plant Cell 10.1105/tpc.111.083337
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.
4
Supplemental Data. Luo et al. (2011). Plant Cell 10.1105/tpc.111.083337
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.
5
Supplemental Data. Luo et al. (2011). Plant Cell 10.1105/tpc.111.083337
Supplemental Table 1. Primers used in this study.
T-D#A insertion
MIPS1
MIPS2
MIPS3
sense
AGA ACA TTG TGC CAC TCC CTG GG
antisense
CCT GAT CGC CAT ATC AAT TAG ACC
sense
CGC TAT GGC TAG AGC TAG AGT TCT
antisense
CGA TAT AGC CAA TTC GAT AAG ACC
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
MIPS2
MIPS3
TUB2
sense
CGT TGG GAT GAA CGA TAC AAT
antisense
ACA TTG TTC CTG ATC GCC ATA
sense
GAT CAT GGA GTA CAA GTG AAG G
antisense
TAA ACC GAC CAG ATC CAT ATC
sense
ATG ATT CTC GAG TAC AAG TGA TC
antisense
CGC GAG ATA CCAT TAC ATA TT
sense
GTT CTC GAT GTT GTT CGT AAG
antisense
TGT AAG GCT CAA CCA CAG TAT
sense
CGA CTT GCA GAA ACA GCT CAG G
antisense
CGC AAC ATA CGG TAC ATA CTT
sense
TCC TTG TTG GAG CTG GCA TT
antisense
AAA CTT CCC CTC TCC TTC AGC
sense
GCT GGT GTT ATC GCC AAT CGT
antisense
CCT CAT CCT TGT CCA AAG ATG A
sense
GTT CTC GAT GTT GTT CGT AAG
antisense
TGT AAG GCT CAA CCA CAG TAT
sense
CCA GCA CAT CAA CAT CAC CCT
antisense
CTC AAT AAA CAT TTT TGC AGA G
sense
TGT CGA TGC GGG ACG TTG AAC
antisense
CTC GAT GAA CAT TTT CTC TGG
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
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
RT PCR
MIPS1
MIPS2
MIPS3
TUB2
Promoter clone
MIPS1
MIPS2
MIPS3
CDS clone
MIPS1
6
Supplemental Data. Luo et al. (2011). Plant Cell 10.1105/tpc.111.083337
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
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
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
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
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
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
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
MIPS3
INO1
Over expression
PIS1
PIS2
UGT84B1
OX
lines
insertion
T-D#A
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.
7
Supplemental Data. Luo et al. (2011). Plant Cell 10.1105/tpc.111.083337
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
8
Supplemental Data. Luo et al. (2011). Plant Cell 10.1105/tpc.111.083337
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
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Supplemental Data. Luo et al. (2011). Plant Cell 10.1105/tpc.111.083337
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
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Supplemental Data. Luo et al. (2011). Plant Cell 10.1105/tpc.111.083337
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
11