supplementary figure 1. ca19-9 and sle x expression correlated with fut3 and fut6

5
Supplementary Figure 1. CA19-9 and sLe x expression correlated with FUT3 and FUT6 A C 95.3% 7.6% 0.8% 26.3% HT29 HCT15 Colo205 Ls174T FITC Cell number Isotypic control CA19-9 B concentration (IU/ml/10 6 cells) Relative expression ( / ß-actin) FUT3 FUT6 Cell number FITC sLe x 99.83% 50.2% 0.7% LS180 42.2% Isotypic control 57.4% 88.7% HT29 HCT15 Colo205 Ls174T LS180

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B. A. HT29. HCT15. Colo205. Ls174T. LS180. Isotypic control. concentration (IU/ml/10 6 cells). 7.6%. 0.8%. 95.3%. 26.3%. 42.2%. CA19-9. Cell number. FITC. LS180. FUT3. FUT6. HT29. HCT15. Colo205. Ls174T. C. Isotypic control. Relative expression ( / ß-actin). 50.2%. - PowerPoint PPT Presentation

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Supplementary Figure 1. CA19-9 and sLex expression correlated with FUT3 and FUT6

A

C

95.3%7.6% 0.8% 26.3%

HT29 HCT15 Colo205 Ls174T

FITC

Cel

l nu

mb

er

Isotypic control

CA19-9

B

con

cen

trat

ion

(IU

/ml/1

06 c

ells

)R

elat

ive

exp

ress

ion

( /

ß-a

ctin

) FUT3 FUT6

Cel

l nu

mb

er

FITC

sLex99.83%50.2% 0.7%

LS180

42.2%

Isotypic control

57.4% 88.7%

HT29 HCT15 Colo205 Ls174T LS180

A B

FUT3

LS174TColo205 No

treat

Rando

m

ß-actin

FUT6

siFUT3

siFUT3

+ siF

UT6

No tre

at

Rando

m

siFUT6

siFUT3

+ siF

UT6

ß-actin

Supplementary Figure 2. Suppression of FUT3 and/or FUT6 by siRNAs

1.0 0.94 0.23 0.22

1.0 1.02 0.28 0.30

FUT5

FUT1

FUT2

FUT4

FUT7

FUT8

random

siRNA1

siRNA2

Supplementary Figure 3. Suppression of FUT3 and FUT6 inhibits Smad2 phosphorylation upon TGF-ß stimulation

siFUT5siFUT1 siFUT2 siFUT4 siFUT7 siFUT8NT Random

TGF-β – –+ + – + – + – + – –+ + – +

pSmad2

Smad2

A

B

1.0 0.26 0.28

1.0 0.36 0.32

1.0 0.21 0.22

1.0 0.44 0.72

1.0 0.33 0.43

1.0 0.27 0.35

1.0 1.36 1.0 1.25 1.0 3.17 1.0 1.38 1.0 1.33 1.0 1.42 1.0 1.52 1.0 1.35

Supplementary Figure 4. Fucosylation of TßR-I was not affected by TGF-ß stimulation

IgG 0 6 12

TGF-β (hr)

TßR-I IP/ AAL IB

TßR-I

TßR-I IP/ TßRI IB TßR-I

1.00 1.05 1.04

Supplementary Figure 5. TßRs bound-CA19-9 and sLex inhibited by siFUT3 and/or siFUT6

Rando

m

siFUT3

siFUT6

NT siFUT3

+ 6

TßR-I IP/ CA19-9 IB

1.00 1.05 0.35 0.25 0.29

Rando

m

siFUT3

siFUT6

NT siFUT3

+ 6

TßR-I IP/ sLex IB

1.00 0.93 0.34 0.23 0.16