Post on 06-Mar-2018
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