uab barcelona · 927
TRANSCRIPT
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A B
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2b 1
2a 3
4
4S
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A B
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A B
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21 Gy
95 days 4.5 months 6 months
Busulfan Melphalan
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α
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780
13 min
200000
Time after D-luciferin Addition (min)
BLI
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Cell number
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1×1007
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∼
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0 1000 2000 3000 4000 5000 60000
25
50
75
100
High (Above Median)Low (Below median)
n=125
n=126
p= 1.76·10-9
LFG
Days
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0 1000 2000 3000 4000 5000 60000
25
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High (Above Median)Low (Below median)
n=125
n=126
p=0.024
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Days
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0 1000 2000 3000 4000 5000 60000
25
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n=125
n=126
p= 5.6·10-4
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0 1000 2000 3000 4000 5000 60000
10
20
30
40
50
60
70
80
90
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n=124
n=94
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Follow up in days
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n=51
n=51
p= 4.7 e-4 p= 0.232
n=36
n=36
High (Above median) Low (Below median)
GSE16476
B C
![Page 116: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/116.jpg)
LFG
Favorable Unfavorable0.01
0.1
1
10
100
**
Rel
ativ
e m
RN
A ex
pres
sion
![Page 117: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/117.jpg)
A8
B8
D4
D12
MY
CN
non
-am
plifi
ed, f
avor
able
his
tolo
gy
MY
CN
am
plifi
ed, u
nfav
orab
le h
isto
logy
Magnification
![Page 118: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/118.jpg)
α
SK-N-BE(2)
RA (days) 0 1 3 5
LFG
α-tubulin
1.00 1.91 2.04 4.07
35
50
kDa
SH-SY5Y
RA (days) 0 1 3 5
LFG
α-tubulin
1.00 2.36 13.08 9.03
35
50
kDa
SH-SY5Y
0 1 2 3 4 5 60
1
2
3
4
0
50
100
150
200LFGTRKB
Retinoic Acid (days)
Rel
ativ
e LF
G e
xpre
ssio
n
Relative TR
KB expression
SK-N-BE(2)
0 1 2 3 4 5 60
1
2
3
0
20
40
60
80
100LFGTRKB
Retinoic Acid (days)
Rel
ativ
e LF
G e
xpre
ssio
n
Relative TR
KB expression
A
B
![Page 119: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/119.jpg)
Adhere
nt
Sphere
Adhere
nt
Sphere
Adhere
nt
Sphere
0.0
0.5
1.0
1.5SK-N-BE(2)IMR-5IMR-32
***
LFG
Rel
ativ
e m
RN
A ex
pres
sion
Adherent Sphere
IMR-5 A B
![Page 120: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/120.jpg)
α
SH-SY5Y +
Scr1 LFG1 Scr2 LFG2
LFG
α-tubulin
1 1 0.37 0.40
CHLA-90 + shRNA
Scr1 LFG1 Scr2 LFG2
LFG
α-tubulin
1 1 0.35 0.83
35
50
35
50
kDa kDa
A B
![Page 121: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/121.jpg)
SH-SY5Y
0 1 2 3 4 5 6 70
1
2
3
4 shScrshLFGshScr 2shLFG 2
** ##,
Days
Cel
l Pro
lifer
atio
n (A
bs 5
90nm
)CHLA-90
0 1 2 3 4 5 6 70
1
2
3
4 shScrshLFGshScr 2shLFG 2
** #,
Days
Cel
l Pro
lifer
atio
n (A
bs 5
90nm
)
A B
CHLA-90
0 30 60 900.0
0.5
1.0
1.5
2.0 shScrshLFG
**
Time (min)
Cel
l Adh
esio
n (A
bs 5
90nm
)
SH-SY5Y
0 30 60 90 1200.0
0.1
0.2
0.3
0.4
0.5 shScrshLFG
*
Time (min)
Cel
l Adh
esio
n (A
bs 5
90nm
)
A B
![Page 122: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/122.jpg)
shScr shLFG
LFG
α-tubulin
SK-N-BE(2)
1 0.31
35
50
kDa shScr shLFG
LFG
α-tubulin
IMR-32
1 0.63
35
50
kDa
A
B
C SK-N-BE(2)
0 20 40 60 80 1000.0
0.2
0.4
0.6 shScrshLFG
Time (min)
Cel
l Adh
esio
n (A
bs 5
90nm
)
IMR-32
0 20 40 60 80 1000.0
0.5
1.0
1.5
2.0 shScrshLFG
Time (min)
Cel
l Adh
esio
n (A
bs 5
90nm
)SK-N-BE(2)
0 1 2 3 40.0
0.5
1.0
1.5
2.0 shScrshLFG
Time (days)
Cel
l Pro
lifer
atio
n (A
bs 5
90nm
)
IMR-32
0 1 2 3 40
1
2
3
4
5 shLFGshScr
Time (days)
Cel
l Pro
lifer
atio
n (A
bs 5
90nm
)
![Page 123: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/123.jpg)
shScr shLFG A B
shScr shLFG0
5
10
15 **
% o
f sph
ere-
form
ing
cells
![Page 124: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/124.jpg)
ABCG2 CD133 KLF4 NANOG NESTIN NOTCH3 OCT4 SOX2
1
2 shScrshLFG
****Fo
ld c
hang
e vs
shS
cr
SNAI1 TCF3 ZEB1 ERBB3 KRT19 SNAI2 TWIST1
1
2 shScrshLFG ***** ***
Fold
cha
nge
vs s
hScr
![Page 125: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/125.jpg)
TWIST1
High LFG Low LFG0
250
500
750p<0.0001
mR
NA
expr
essi
on (a
.u.)
FN1
High LFG Low LFG0
500
1000
1500
2000p=0.0041
mR
NA
expr
essi
on (a
.u.)
SNAI2
High LFG Low LFG0
25
50
75p=0.0060
mR
NA
expr
essi
on (a
.u.)
SPARC
High LFG Low LFG0
500
1000
1500
2000
2500p=0.0370
mR
NA
expr
essi
on (a
.u.)
TCF3
High LFG Low LFG0
50
100
150
200p<0.0001
mR
NA
expr
essi
on (a
.u.)
CALD1
High LFG Low LFG0
200
400
600p=0.0248
mR
NA
expr
essi
on (a
.u.)
ERBB3
High LFG Low LFG0
10
20
30
40p=0.0004
mR
NA
expr
essi
on (a
.u.)
KRT19
High LFG Low LFG0
200
400
600
800
1000
1200p=0.0163
mR
NA
expr
essi
on (a
.u.)
A
B
![Page 126: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/126.jpg)
shScr shLFG0
2
4
6
*
Mig
ratio
n (fo
ld-in
duct
ion)
![Page 127: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/127.jpg)
shScr shLFG0
50
100
150
Cel
l num
ber p
er fi
eld
A B
![Page 128: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/128.jpg)
0 50000 100000 150000 2000000
2×1007
4×1007
6×1007
8×1007SH-SY5Y-shScrSH-SY5Y-shLFG
r2=0.99, p<0.0001
r2=0.99, p<0.0001
Number of Cells/Well
BLI I
nten
sity
(ph/
s)
![Page 129: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/129.jpg)
8 11 15 170
20
40
60
80
100 shScrshLFG
Days post-injection
Tum
or in
cide
nce
(%)
0 5 10 15 20 250
500
1000
1500shScrshLFG
Days post-injection
Tum
or v
olum
e (m
m3 )
shScr shLFG0.0
0.5
1.0
1.5
Tum
or w
eigh
t (g)
A B
![Page 130: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/130.jpg)
B
1.5
1.0
0.5
x106
Radiance (p/sec/cm2/sr)
shSc
r sh
LFG
A
1.5
1.0
0.5
x106
Radiance (p/sec/cm2/sr)
2.0
shSc
r sh
LFG
shScr shLFG105
106
107
108
109
Tota
l BLI
(ph/
sec)
shScr shLFG104
105
106
107
108
Tota
l BLI
( ph/
sec)
![Page 131: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/131.jpg)
B
shLF
G
shSc
r
A
shLF
G
shSc
r
shScr shLFG0
5
10
15 *
Num
ber o
f Met
asta
ses
shScr shLFG0
20
40
60
Num
ber o
f Met
asta
ses
![Page 132: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/132.jpg)
PC1 60.5%
PC2
29.6
%
0
010
5 10- 5- 10
515
-10
-15
-5
shLFG1
CTL1
CTL2
CTL3
shLFG2
shLFG3
PC1 60.5%
PC2
29.6
%
0
010
5 10- 5- 10
515
-10
-15
-5
shLFG1
CTL1
CTL2
CTL3
shLFG2
shLFG3
![Page 133: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/133.jpg)
Colorec
tal ca
ncer
Cell cy
cle
Pancre
atic c
ance
r
Adhere
ns ju
nctio
n
Cell ad
hesio
n mole
cules
Osteoc
last d
ifferen
tiatio
n
Focal
adhe
sion
Pathway
s in c
ance
r
Metabo
lic pa
thway
s0
1
2
3
4
Odd
s R
atio
![Page 134: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/134.jpg)
FGF14 STK36 BCL2 RAP1A TJP1 ITGA9 LFG-3
-2
-1
1
2
3 ArrayqPCR
Fold
cha
nge
vs s
hScr
Focal adhesion Cell adhesion
Adherent junctions
Cell cyclePathways in Cancer
![Page 135: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/135.jpg)
Tet21N
Untreated Doxycycline
![Page 136: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/136.jpg)
α
α
Tet21N + Dox (days)
0 1 3 5
LFG
α-tubulin
7
MYCN
1.00 0.55 1.76 3.66 4.53
Tet21N
0 1 2 3 4 5 6 70
10
20
30
0.0
0.5
1.0
1.5LFGMYCN
Dox (days)
Rel
ativ
e LF
G e
xpre
ssio
n
Relative M
YCN
expression
A B
35
50
64
kDa
IMR-32 shMYCN
0 1 2 30.0
0.5
1.0
1.5
2.0
2.5
0.0
0.5
1.0
1.5LFGMYCN
Dox (days)
Rel
ativ
e LF
G e
xpre
ssio
nR
elative MYC
N expression
![Page 137: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/137.jpg)
1 2 30.0
0.5
1.0
1.5
2.0 VehicleDox
Days
LFG
mR
NA
expr
essi
on (a
.u.)
![Page 138: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/138.jpg)
+ D
ox
- Dox
LFG
![Page 139: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/139.jpg)
LAI-5S
0 1 2 30
5
10
15 5-azaDMSO
days
Rel
ativ
e LF
G e
xpre
ssio
n
SK-N-BE(2)
0 1 2 30
1
2
3
4
5 5-azaDMSO
days
Rel
ativ
e LF
G e
xpre
ssio
n
Tet21N
0 1 2 30
10
20
30 5-azaDMSO
days
Rel
ativ
e LF
G e
xpre
ssio
n
LAI-5S
0 1 20
2
4
6
8
10 NaBPBS
daysR
elat
ive
LFG
exp
ress
ion
SK-N-BE(2)
0 1 20
1
2
3 NaBPBS
days
Rel
ativ
e LF
G e
xpre
ssio
n
Tet21N
0 1 20
20
40
60
80
100 NaBPBS
days
Rel
ativ
e LF
G e
xpre
ssio
n
A
B
![Page 140: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/140.jpg)
![Page 141: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/141.jpg)
![Page 142: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/142.jpg)
![Page 143: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/143.jpg)
∼
![Page 144: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/144.jpg)
![Page 145: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/145.jpg)
![Page 146: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/146.jpg)
![Page 147: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/147.jpg)
![Page 148: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/148.jpg)
![Page 149: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/149.jpg)
![Page 150: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/150.jpg)
![Page 151: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/151.jpg)
![Page 152: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/152.jpg)
![Page 153: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/153.jpg)
![Page 154: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/154.jpg)
![Page 155: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/155.jpg)
![Page 156: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/156.jpg)
![Page 157: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/157.jpg)
![Page 158: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/158.jpg)
![Page 159: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/159.jpg)
![Page 160: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/160.jpg)
![Page 161: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/161.jpg)
![Page 162: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/162.jpg)
![Page 163: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/163.jpg)
![Page 164: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/164.jpg)
![Page 165: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/165.jpg)
![Page 166: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/166.jpg)
![Page 167: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/167.jpg)
![Page 168: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/168.jpg)
![Page 169: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/169.jpg)
![Page 170: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/170.jpg)
![Page 171: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/171.jpg)
![Page 172: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/172.jpg)
![Page 173: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/173.jpg)
![Page 174: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/174.jpg)
![Page 175: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/175.jpg)
![Page 176: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/176.jpg)
![Page 177: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/177.jpg)
![Page 178: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/178.jpg)
![Page 179: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/179.jpg)
![Page 180: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/180.jpg)
![Page 181: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/181.jpg)
![Page 182: UAB Barcelona · 927](https://reader030.vdocuments.net/reader030/viewer/2022013023/5fb28f52b40e0d0d890a4630/html5/thumbnails/182.jpg)
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