Figure S1. A...Figure S1. 250 150 100 75 A kDa 50 37 25 Heavy chain Light chain 20 15 2 2.5 3 2 2.5...

8
Figure S1. 250 150 100 75 A kDa 50 37 25 Heavy chain Light chain 20 15 2 2.5 3 2 2.5 3 Control IgG + 0.5μg mTM4SF5R2-3 Anti-TM4SF5 Ab + 1μg mTM4SF5R2-3 + 5 μg mTM4SF5R2-3 Control IgG + 0.5 μg hTM4SF5R2-3 Anti-TM4SF5 Ab + 1 μg hTM4SF5R2-3 + 5 μg hTM4SF5R2-3 hTM4SF5R2-3 peptide mTM4SF5R2-3 peptide B C 0 0.5 1 1.5 2 A450nm 0 0.5 1 1.5 2 A450nm + 10 μg mTM4SF5R2-3 + 10 μg hTM4SF5R2-3 0 0 Anti-TM4SF5 Ab (ng/ml) Anti-TM4SF5 Ab (ng/ml)

Transcript of Figure S1. A...Figure S1. 250 150 100 75 A kDa 50 37 25 Heavy chain Light chain 20 15 2 2.5 3 2 2.5...

Figure S1.

250150100

75

AkDa

50

37

25

Heavy chain

Light chain

20

15

2

2.5

3

2

2.5

3Control IgG

+ 0.5μg mTM4SF5R2-3

Anti-TM4SF5 Ab

+ 1μg mTM4SF5R2-3

+ 5 μg mTM4SF5R2-3

Control IgG

+ 0.5 μg hTM4SF5R2-3

Anti-TM4SF5 Ab

+ 1 μg hTM4SF5R2-3

+ 5 μg hTM4SF5R2-3

hTM4SF5R2-3 peptide mTM4SF5R2-3 peptideB C

0

0.5

1

1.5

2

A45

0nm

0

0.5

1

1.5

2

A45

0nm

μ+ 10 μg mTM4SF5R2-3 + 10 μg hTM4SF5R2-3

0 0

Anti-TM4SF5 Ab (ng/ml)Anti-TM4SF5 Ab (ng/ml)

TM4SF superfamily expression B cell epitope sequences of TM4SF superfamily A B

Figure S2.

Name Fold

TM4SF4 37.369553

TM4SF5 27.422548

TM4SF1 6.783213

TM4SF18 0 401847

Name Sequences

TM4SF1 DTSTWSECTEPKHI

TM4SF4 DEALWNKCREPLNV

TM4SF5 NRTLWDRCEAPPRV

TM4SF18 DSSIWIQCLEPAHV

(microarray data)Huh-7 / SNU-739

corresponding to TM4SF5

2.5 2.5peptide coating peptide coating

TM4SF1

TM4SF18 0.401847

TM4SF19 0.17190197

TM4SF18 DSSIWIQCLEPAHV

TM4SF19 DRSLWNSVCLEPSAA

TM4SF1

Anti-TM4SF5 AbC Normal IgG

0 5

1

1.5

2

0 5

1

1.5

2

2.5

A45

0nm

A45

0nm

TM4SF1TM4SF4TM4SF5TM4SF18TM4SF19

TM4SF1TM4SF4TM4SF5TM4SF18TM4SF19

0

0.5

0

0.5

Pancreas Ileum Oeoum Kidney cortex Kidney medulla

Figure S3.

Colon Rectum Gall bladder Adrenal gland Skin

C b l t C b ll B l li P t t T tiCerebral cortex Cerebellum Basal ganglia Prostate Testis

Hippocampus Heart Spleen Exocervix Endocervix

Lymph node Thymus Tongue Pro-endometrium Sec-endometrium

Skeletal muscle Tonsil Parotid gland Myometrium Pro-endometrium

Ureter Soft tissue Lung Ovary Fallopian tube

Liver Breast Bronchus Placenta amnion Placenta chorionvilli

Esophagus Stomach Spinal cord Placenta basal plate Eye

A204 A204(II) A204(III)B CA

Figure S4.

No. Sex Age E-S grade Ab staining1 F 48 I +++2 M 55 I ++3 M 51 II ++4 M 49 III ++5 M 42 III +++/++++6 M 40 II +++

No. Sex Age E-S grade Ab staining1 F 65 IV ++2 M 53 III +++3 M 41 III +++4 M 49 I to II ++++5 M 57 I and III +++6 M 40 III +++

No. Sex Age E-S grade Ab staining1 F 59 I to II +++2 M 71 II to III ++3 M 67 II to III ++4 M 67 II to III +++5 F 59 I to II +++/++++6 M 82 I to II ++++

7 M 50 II +++8 M 45 II ++9 M 43 III +++

10 M 42 III ++++11 F 50 II +++12 M 53 II +++13 M 55 II +++

7 M 46 I to II ++++8 M 39 III ++++9 M 72 II to III +++

10 M 46 III ++++11 M 42 - ++12 M 58 III +++13 M 52 I to II ++++

7 M 11 - ++8 F 37 II +++9 M 77 III +++

10 F 64 II to III +++11 M 60 - ++++12 M 63 III ++++13 M 45 II to III ++++

14 M 49 III to IV ++++15 M 40 IV +++16 M 51 II +++17 M 56 II ++++18 M 33 II ++19 F 56 IV ++20 M 63 II +++

14 M 54 II to IV ++++15 M 53 - +++16 M 58 II to III ++17 F 39 III ++18 M 48 I to II ++++19 M 67 I to II ++++20 M 36 II to III +++

14 F 48 II to III +++15 M 64 I, II, III ++16 M 58 I, II, III +++17 M 50 II to III ++++18 M 55 II +++19 M 62 I +++20 M 57 II, III +++20 M 63 II +++

21 M 52 II +++22 M 41 III ++23 M 60 III +++24 M 66 II +++25 M 70 II ++++26 M 37 III ++++27 M 51 III ++

20 M 36 II to III +++21 F 39 II to III ++22 M 55 III ++++23 F 42 I to II +++24 M 52 II +++25 M 61 II to III +++26 M 55 I to II ++++27 M 61 III ++

20 M 57 II, III21 M 47 III +++22 M 59 III ++23 F 67 III +++/++24 F 65 II+III ++++25 M 68 I to II +++26 F 52 II to III ++27 M 56 II ++27 M 51 III ++

28 M 60 III +++29 M 47 II ++++30 M 70 II to III ++++31 M 63 III ++32 M 56 III +++33 M 56 III +++34 M 72 II t III ++++

27 M 61 III ++28 M 50 II +++29 F 60 I ++30 M 50 III ++++31 M 51 II +++32 M 56 I to II +++33 F 32 II +++34 M 57 III ++++

27 M 56 II ++28 M 49 III ++29 M 42 III +++30 M 67 III +++31 M 67 - ++32 M 74 I ++++33 M 68 II ++++34 M 56 II to III ++++34 M 72 II to III ++++

35 M 70 III ++++

HCC > 75% ++++

HCC 50-74% +++

HCC 11-49% ++

34 M 57 III ++++35 M 47 II +++

34 M 56 II to III ++++35 M 47 II ++

Adrenal grandPheochromolytoma

EsophagusSCC

Salivary grandPleomorphic adenoma

IleumLarge cell lymphoma

ThyroidPapillary CA

Figure S5.

Uriary bladderPapillary urothelical CA

Soft tissueLymphoma

SkinSCC

TonsilSCC

LungSCC

PancreasAdenoca

LungAdeno CA

ProstateAdenoca

StomachAdenoca

StomachSignet ring cell CA

Paratoid glandPleomorphic adenoma

KidneyRCC

KidneyRCC

OvaryYolk sac tumor

UterusAdenoca

UterusSCC

BreastInvasive lobular carcinoma

CecumAdenoca

TestisSeminoma

ThymusThymoma

A

Figure S6.

E-cadherin Nuclei Merged E-cadherin Nuclei Merged

PB

S

A B

PB

S

Huh-7 SNU-739

IgG

2a

b

IgG

2a

b

Ant

i-TM

4SF

5 A

b

Ant

i-TM

4SF

5 A

b

(-) E-cadherin Nuclei Merged (-) E-cadherin Nuclei Merged

b Ab

IgG2a Anti-TM4SF5 AbC IgG2a Anti-TM4SF5 Ab

Ant

i-TM

4SF

5 A

b

Ant

i-TM

4SF

5 A

0 1 2 3 4 (days) 0 1 2 3 4 (days) 0 1 2 3 4 (days) 0 1 2 3 4 (days)

Huh

-7

SN

U-7

39E-cadherin

β-actin

E-cadherin

β-actin

Figure S7.

PBS IgG2a Anti-TM4SF5 Ab PBS IgG2a Anti-TM4SF5 AbA B

g g

Huh

-7

Huh

-7

SN

U-7

39

SN

U-7

39

CNormal IgG Anti-TM4SF5 Ab

1

Huh

-7 p27kip1

RhoA

0.5 1 30 (h)6 0.5 3 6

(h)

Normal IgG Anti-TM4SF5 Ab

Rho

A in

tera

ctio

n (%

)

50

100

150

50

100

150***Normal IgGAnti-TM4SF5 Ab

D

SN

U-7

39 p27kip1

RhoA

(h)10.5 1 30 6 0.5 3 6

P Normal IgG Anti-TM4SF5 Ab

PγS

*

0.5 1 3 6 (h) 0.5 1 3 6

Huh-7 cells SNU-739 cells

p27k

ip1

/ R

0

50

0

50

(h)

DP

TPγS Normal IgG Anti-TM4SF5 Ab

0.5 1 3 0.5 1 30GD

(h)

Huh

-7 GTP-RhoA

Total RhoA

GT

P

Normal IgGAnti-TM4SF5 Ab

Rho

A a

ctiv

ity (

%)

50

100

150

200

250

50

100

150

200

250 *

0.5 1 3 0.5 1 30GD

GT

(h)

SN

U-7

39 GTP-RhoA

Total RhoA

0.5 1 3 0.5 1 3(h)

Huh-7 cells SNU-739 cells

R

0

50

0

50

(h)

Figure S8.

Untreated 0 h 3 h 6 h

12 h 24 h 48 h 96 h