96 well plateでのHigh-throughput...
Transcript of 96 well plateでのHigh-throughput...
96 well plateでのHigh-throughput Screening
RNAi効果に影響を与える遺伝子群のスクリーニング
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<Cell density> 1 x 105 cells/ml → 170 µl/well <Transfection> Opti-MEM 10 µl pGL3-Cont 150 nl pRL-SV40 15 ng shRNA + Opti-MEM 10 µl FL2000 0.4 µl Medium Change 50 µl <Assay> PLB=30 µl 3回の実験で、 誤差を確認。 (HeLa)
GN.NAB.4
0
5
10
15
20
25
30
0 50 100 150 200 250 300 350 400
NAB.4-R1
NAB.4-R2
NAB.4-R3
binding.07
NAB.3
NTB.7
TF.4
TF.5
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0
10
20
30
40
50
0 50 100 150 200 250 300 350 400
U575
U588
U753
H95
H129
Distribution of Renilla luciferase activity (each experiment)
Renilla luciferase activity (%)
Sam
ple
No.
Sam
ple
No.
Sam
ple
No.
Distribution of Renilla luciferase activity (total)
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Renilla luciferase activity (%)
Sam
ple
No.
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Renilla luciferase activity (%)
Sam
ple
No. Luc Assay (379)-10
50 150
GN.NAB.4
0
20
40
60
80
100
120
ABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGH 1 2 3 4 5 6 7 8 9 10 11 12
Rela
tive
luc
activ
ity (%
)
Control pSUPER-FL774 pSUPERretroPuro
EIF2C2
1000
10000
100000Renilla
luc
activ
ity
EIF3S6IP
EIF5
ANKHD1
HBS1L
DBF4
BOLL
CPEB1
ZNF385 EEFSEC
BRF2 UPF2 EIF2C1 SYNJ2
060908-15
0
20
40
60
80
100
120
1000
10000
100000
Control pSUPER-FL774 pSUPERretroPuro
ABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCD
1 2 3 4 5 6 7 8 9 10 11 12
GN.NAB.3 Re
lativ
e lu
c ac
tivity
(%)
Renilla
luc
activ
ity
PUM2
EXOSC8 POLR2G
RBM3 SFRS14
RBM15 �
METTL3
MGC4562
MOV10L1
MSI1
NOL5A
NSUN6
NUFIP1
NXF1
NXF3 OAS2
OAS3
SF3B14
PABPC5
PAPOLA
PNPT1
PSMA1 PUM1
RBMS3 RDBP
RNASE1 RNASEH2A
RNASET2 RPL4
RPN1
RPS21
RPS4Y2 SF3A2
SFRS3 SKIV2L
SNRPA1
060926
0
20
40
60
80
100
120
����
�����
������
Control pSUPER-FL774 pSUPERretroPuro
ABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCD
1 2 3 4 5 6 7 8 9 10 11 12
GN.NTB.7 Re
lativ
e lu
c ac
tivity
(%)
Renilla
luc
activ
ity
ROCK1
PRKG1
PSKH2
PSMC1
PSMC6
PTK2
PTK6
PTK7 RET
ARHGAP17
ROCK2
ROS1
RPS6KA1
RPS6KB2 SGK2
CLPB SMC1A SMC5 SPATA5
PRKCD
PRKCG
PRKCH
PKN1
PKN2
PRKD3
PRKCQ
WNK1
WNK2
WNK4 ATAD2
RAD50 RARSL
RPS6KA6
SNF1LK SRMS
SRPK2 STK10
STK16
STK17A
STK19 TSSK3
060929
PRKG2
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Control pSUPER-FL774 pSUPERretroPuro
ABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCD
1 2 3 4 5 6 7 8 9 10 11 12 � � � �
�����
������
GN.TF.4 Re
lativ
e lu
c ac
tivity
(%)
Renilla
luc
activ
ity
Rb1 NR2E1
NR2F1
PKNOX1
PPARG
RAX
SATB1
NR2F2
NR5A1
GATAD2A
GATAD2B
PBX1
PBX2 PGR
JARID1A REL
RERE
POU6F2 RUNX1 RUNX2
SCMH1
SCML2
060929
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������
Control pSUPER-FL774 pSUPERretroPuro
ABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHEFGH
1 2 3 4 5 6 7 8 9 10 11 12
GN.TF.5 Re
lativ
e lu
c ac
tivity
(%)
Renilla
luc
activ
ity
p53
SOX6
SRF
STAT4
TBX6
TCEA1 TBX4
060929
SLC38A6
SNAPC5
SOX2
SOX5
SPIC
SUPT6H
T
TADA3L TAF13 TAF5L TBX1
TBX5 TCF3
MLX
TEAD4
TFAP2C
THRA
TNRC4
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1000
10000
100000
Rbbp6
GN.binding.07 Re
lativ
e lu
c ac
tivity
(%)
Renilla
luc
activ
ity
Control pSUPER-FL774 pSUPERretroPuro
ABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCD
1 2 3 4 5 6 7 8 9 10 11 12
LRP6
LRPAP1 SMAD7 NMNAT1
PGLYRP3
KBTBD10
SLC9A3R2 SGTA SENP2 SOCS5
SORCS2
CDC42SE1 SPOP
PEA15
GIPC1
LMNA LTBP4
RPE65 MPP6
RTKN
060926
0
20
40
60
80
100
120
1000
10000
100000
GN.binding.07-2 Re
lativ
e lu
c ac
tivity
(%)
Renilla
luc
activ
ity Control
pSUPER-retro-puro EIF2C2
ABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCD
1 2 3 4 5 6 7 8 9 10 11 12
Rbbp6 LRP6 LRPAP1
SMAD7 MPP2 NMNAT1
PGLYRP3
SLC9A3R2 SOCS5
SORCS2 SPOP
MPP6
NPHP1
NUDT5
OIP5 PHIP RCBTB1
RCN2
GIPC1
RIN1
SENP2
RPE65
SLIT3 SMO
SNCAIP
CDC42SE1
TCEB3
061013
0
20
40
60
80
100
120GN.binding.07-1GN.binding.07-2
1000
10000
100000
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!"#$%&'%&(#)*+-
Rbbp6
GN.binding.07-1&2 Re
lativ
e lu
c ac
tivity
(%)
Renilla
luc
activ
ity
Control pSUPER-FL774 pSUPERretroPuro
ABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCD
1 2 3 4 5 6 7 8 9 10 11 12
LRP6
LRPAP1 SMAD7 NMNAT1
PGLYRP3
KBTBD10
SLC9A3R2 SGTA SENP2 SOCS5
SORCS2
CDC42SE1 SPOP
PEA15
GIPC1
LMNA LTBP4
RPE65 MPP6
RTKN
0
20
40
60
80
100
120
1000
10000
100000
GN.NTB.2 Re
lativ
e lu
c ac
tivity
(%)
Renilla
luc
activ
ity Control
pSUPER-retro-puro EIF2C2
ABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCD
1 2 3 4 5 6 7 8 9 10 11 12
CAMK1G CASK
CDK2
CDK4
CDKL2
CDKL5 YARS2 CLK3
DDR2 BMP2K
TRIB1 CACNA1B
BMPR1B
BMPR2
BUB1 NUBPL
TSSK4
CAMK1
CAMK2A CCT7
CDC7 CLK1 CLK4
CSNK1E
CSNK1G3
061013
CDC2
0
20
40
60
80
100
120
1000
10000
100000
GN.DNAB.1 Re
lativ
e lu
c ac
tivity
(%)
Renilla
luc
activ
ity Control
pSUPER-retro-puro EIF2C2
ABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCD
1 2 3 4 5 6 7 8 9 10 11 12
SUV39H1
POLH POLQ
CHAF1A
RBMS1 IFI16
SAFB
JTV1
ERCC1 MSH3
MSH5
NBN
RAD1
RAD18 REV1L
XPA XRCC2
CENPA
ING2 CBX2
NCOA6
ARID4B MLH3
061013
0
20
40
60
80
100
120
1000
10000
100000
GN.NAB.2 Re
lativ
e lu
c ac
tivity
(%)
Renilla
luc
activ
ity Control
pSUPER-retro-puro EIF2C2
ABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGH
1 2 3 4 5 6 7 8 9 10 11 12
A2BP1
ADAT1 CSTF3
CUGBP2
HNRPA2B1
KIAA0427
LGTN
DDX21 DDX43
EIF2S1 CIRBP
BAT1
061017
EIF4G1
DAZL EIF4G3
EIF4E2
0
20
40
60
80
100
120
STAU1
1000
10000
100000
GN.NAB.1 Re
lativ
e lu
c ac
tivity
(%)
Renilla
luc
activ
ity Control
pSUPER-retro-puro EIF2C2
ABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCD 1 2 3 4 5 6 7 8 9 10 11 12
WDR57 KHSRP
LSM8 LSMD1
SF3B1
SNRPB
SFRS11
SNRPN
EFTUD2
ELAVL4
ADARB2
CARF
DICER1
LRRFIP1
PRKRA TARBP2
SF3B4 LSM11 AUH
CPSF1 ELAVL2
RNASEN (DROSHA)
DUS2L
061017
PABPC3
PABPC4
STAU2
ADAD1
0
20
40
60
80
100
120
1000
10000
100000
GN.Hs_71_TF.1 Re
lativ
e lu
c ac
tivity
(%)
Renilla
luc
activ
ity Control
pSUPER-retro-puro EIF2C2
ABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCD
1 2 3 4 5 6 7 8 9 10 11 12
AHR
ALX4
AR ARNTL
ATF7
ATOH1 BNC1
BRD8
BRF1
BTAF1
NULL
TSHZ2 RUNX1T1
CEBPB
ARNT ARNT2
ARNTL2
ASCL1 ATF3
ATF6 ATRX BACH1
BAPX1
BARX1
BARX2
MKX
CART1
CBFA2T3
CBFB
061017
CDX2
CDX4
0
20
40
60
80
100
120
1000
10000
100000
GN.A_L.08 Re
lativ
e lu
c ac
tivity
(%)
Renilla
luc
activ
ity Control
pSUPER-retro-puro EIF2C2
ABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCD
1 2 3 4 5 6 7 8 9 10 11 12
ALKBH3
PSRC1 DC-UbP
PCBD2
DCDC2
ENY2
C3orf37
DIDO1
CD55
CYP26C1
CYFIP1
CYBRD1
CXorf6 CTLA4 SLC44A2
061027
DCP1B DCPS
DCP2
0
20
40
60
80
100
120
1000
10000
100000
GN.NAB.5 Re
lativ
e lu
c ac
tivity
(%)
Renilla
luc
activ
ity Control
pSUPER-retro-puro EIF2C2
ABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCD
1 2 3 4 5 6 7 8 9 10 11 12
DDX42
DDX51
DDX54 DHX35
ADPGK
ZNF540 DDX59
DRB1
GPATC1
ENDOGL1
ENPP2
ZNF669 ZNF649
RBM35A
RBM21
ZNF419
NULL ZNF582
C3orf33 ZNF641
ZNF548
ZNF597
DDX19A
061027
0
20
40
60
80
100
120
1000
10000
100000
GN.NAB.6 Re
lativ
e lu
c ac
tivity
(%)
Renilla
luc
activ
ity Control
pSUPER-retro-puro EIF2C2
ABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCD
1 2 3 4 5 6 7 8 9 10 11 12
DHX33
DHX40
KIAA1429
EHD2
WDR52
ZC3H14
ZNF669 ZNF649
ZNF566
FLJ20035
ZNF586 RBED1 ZNF671
061027
IFIH1
0
20
40
60
80
100
120
1000
10000
100000
GN.NAB.7 Re
lativ
e lu
c ac
tivity
(%)
Renilla
luc
activ
ity Control
pSUPER-retro-puro EIF2C2
ABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHEFGH
1 2 3 4 5 6 7 8 9 10 11 12
061110
RAVER1
DDX58 RNASE2 RBM24
RSN
SART3
TRSPAP1
SFRS12
SPEN THOC4
TIPARP U1SNRNPBP
USP6
PARP12
ZCCHC2
RNF113B
ZNF227
ZNF354C
ZNF415
ZNF426
ZNF454
ZNF471
0
20
40
60
80
100
120
GN
.NA
B.4
-1 (H
9)
GN
.NA
B.4
-2 (H
9)
GN
.NA
B.4
-3 (H
9)
GN
.bin
din
g.0
7 (E
12
)G
N.N
AB
.3 (E
12
)G
N.N
TB
.7 (E
12
)G
N.T
F.4 (E
12
)G
N.T
F.5 (A
12
)G
N.b
ind
ing
.07
-2 (A
1)
GN
.bin
din
g.0
7-2
(C5
)G
N.b
ind
ing
.07
-2 (C
9)
GN
.bin
din
g.0
7-2
(E1
2)
GN
.NT
B.2
(A1
)G
N.N
TB
.2 (C
5)
GN
.NT
B.2
(C9
)G
N.N
TB
.2 (E
12
)G
N.D
NA
B.1
(A1
)G
N.D
NA
B.1
(C5
)G
N.D
NA
B.1
(C9
)G
N.D
NA
B.1
(E1
2)
GN
.NA
B.2
(E1
)G
N.N
AB
.2 (C
4)
GN
.NA
B.2
(D8
)G
N.N
AB
.2 (A
12
) G
N.N
AB
.1 (A
1)
GN
.NA
B.1
(C5
)G
N.N
AB
.1 (C
9)
GN
.NA
B.1
(E1
2)
GN
.Hs_
71
_T
F.1 (A
1)
GN
.Hs_
71
_T
F.1 (C
4)
GN
.A_
L.0
8 (A
1)
GN
.A_
L.0
8 (C
5)
GN
.A_
L.0
8 (C
9)
GN
.A_
L.0
8 (E
12
)G
N.N
AB
.5 (C
3)
GN
.NA
B.5
(C6
)G
N.N
AB
.5 (C
9)
GN
.NA
B.5
(E1
2)
GN
.NA
B.6
(A1
)G
N.N
AB
.6 (C
5)
GN
.NA
B.6
(B9
)G
N.N
AB
.6 (E
12
)
GN
.NA
B.4
-1 (H
10
)G
N.N
AB
.4 -2
(H1
0)
GN
.NA
B.4
-3 (H
10
)G
N.b
ind
ing
.07
(F1
2)
GN
.NA
B.3
(F1
2)
GN
.NT
B.7
(F1
2)
GN
.TF.4
(F1
2)
GN
.TF.5
(B1
2)
GN
.NA
B.4
-1 (H
11)
GN
.NA
B.4
-2 (H
11)
GN
.NA
B.4
-3 (H
11)
GN
.bin
din
g.0
7 (G
12
)G
N.N
AB
.3 (G
12
)G
N.N
TB
.7 (G
12
)G
N.T
F.4 (G
12
)G
N.T
F.5 (C
12
)G
N.b
ind
ing
.07
-2 (B
1)
GN
.bin
din
g.0
7-2
(D5
)G
N.b
ind
ing
.07
-2 (D
9)
GN
.bin
din
g.0
7-2
(F1
2)
GN
.NT
B.2
(B1
) G
N.N
TB
.2 (D
5)
GN
.NT
B.2
(D9
)G
N.N
TB
.2 (F
12
)G
N.D
NA
B.1
(B1
) G
N.D
NA
B.1
(D5
)G
N.D
NA
B.1
(D9
)G
N.D
NA
B.1
(F1
2)
GN
.NA
B.2
(F1
)G
N.N
AB
.2 (D
4)
GN
.NA
B.2
(E8
)G
N.N
AB
.2 (B
12
)G
N.N
AB
.1 (B
1)
GN
.NA
B.1
(D5
)G
N.N
AB
.1 (D
9)
GN
.NA
B.1
(F1
2)
GN
.Hs_
71
_T
F.1 (B
1)
GN
.Hs_
71
_T
F.1 (D
4)
GN
.A_
L.0
8 (B
1)
GN
.A_
L.0
8 (D
5)
GN
.A_
L.0
8 (D
9)
GN
.A_
L.0
8 (F
12
)G
N.N
AB
.5 (D
3)
GN
.NA
B.5
(D6
)G
N.N
AB
.5 (D
9)
GN
.NA
B.5
(F1
2)
GN
.NA
B.6
(B1
)G
N.N
AB
.6 (D
5)
GN
.NA
B.6
(C9
)G
N.N
AB
.6 (F
12
)
GN
.bin
din
g.0
7-2
(C1
)G
N.b
ind
ing
.07
-2 (E
5)
GN
.bin
din
g.0
7-2
(E9
)G
N.b
ind
ing
.07
-2G
N.N
AB
.2 (C
1)
GN
.NA
B.2
(E5
)G
N.N
AB
.2 (E
9)
GN
.NA
B.2
(G1
2)
GN
.DN
AB
.1 (C
1)
GN
.DN
AB
.1 (E
5)
GN
.DN
AB
.1 (E
9)
GN
.DN
AB
.1 (G
12
)G
N. N
AB
.2 (G
1)
GN
. NA
B.2
(E4
)G
N. N
AB
.2 (F
8)
GN
. NA
B.2
(C1
2)
GN
. NA
B.1
(C1
)G
N. N
AB
.1 (E
5)
GN
. NA
B.1
(E9
)G
N. N
AB
.1 (G
12
)G
N.H
s_7
1_
TF.1
(C1
)G
N.H
s_7
1_
TF.1
(E4
)G
N.A
_L
.08
(C1
) G
N.A
_L
.08
(E5
)G
N.A
_L
.08
(E9
)G
N.A
_L
.08
(G1
2)
GN
.NA
B.5
(E3
)G
N.N
AB
.5 (E
6)
GN
.NA
B.5
(E9
)G
N.N
AB
.5 (G
12
)G
N.N
AB
.6 (C
1)
GN
.NA
B.6
(E5
)G
N.N
AB
.6 (D
9)
GN
.NA
B.6
(G1
2)
control Re
lativ
e lu
c ac
tivity
(%)
Luc. Luc. + pSUPER -FL774
Luc. + pSUPER-FL774 + pSUPER-retro-puro
Luc. + pSUPER-FL774 + EIF2C2
0
20
40
60
80
100
120
GN
.bin
din
g.0
7-2
(A1
)G
N.N
TB
.2 (A
1)
GN
.DN
AB
.1 (A
1)
GN
.NA
B.1
(A1
)G
N.H
s_7
1_
TF.1
(A1
)G
N.A
_L
.08
(A1
) G
N.N
AB
.6 (A
1)
GN
.TF.5
(A1
2)
GN
.NA
B.2
(A1
2)
GN
.NA
B.6
(B9
)G
N.N
AB
.5 (C
3)
GN
.NA
B.2
(C4
)G
N.H
s_7
1_
TF.1
(C4
)G
N.b
ind
ing
.07
-2 (C
5)
GN
.NT
B.2
(C5
)G
N.D
NA
B.1
(C5
)G
N.N
AB
.1 (C
5)
GN
.A_
L.0
8 (C
5)
GN
.NA
B.6
(C5
)G
N.N
AB
.5 (C
6)
GN
.bin
din
g.0
7-2
(C9
)G
N.N
TB
.2 (C
9)
GN
.DN
AB
.1 (C
9)
GN
.NA
B.1
(C9
)G
N.A
_L
.08
(C9
)G
N.N
AB
.5 (C
9)
GN
.NA
B.2
(D8
)G
N.N
AB
.2 (E
1)
GN
.bin
din
g.0
7 (E
12
)G
N.N
AB
.3 (E
12
)G
N.N
TB
.7 (E
12
)G
N.T
F.4 (E
12
)G
N.b
ind
ing
.07
-2 (E
12
)G
N.N
TB
.2 (E
12
)G
N.D
NA
B.1
(E1
2)
GN
.NA
B.1
(E1
2)
GN
.A_
L.0
8 (E
12
)G
N.N
AB
.5 (E
12
)G
N.N
AB
.6 (E
12
)G
N.N
AB
.4 -1
(H9
)G
N.N
AB
.4 -2
(H9
)G
N.N
AB
.4 -3
(H9
)
GN
.TF.5
(B1
2)
GN
.bin
din
g.0
7 (F
12
)G
N.N
AB
.3 (F
12
)G
N.N
TB
.7 (F
12
)G
N.T
F.4 (F
12
)G
N.N
AB
.4 -1
(H1
0)
GN
.NA
B.4
-2 (H
10
)G
N.N
AB
.4 -3
(H1
0)
GN
.bin
din
g.0
7-2
(B1
)G
N.N
TB
.2 (B
1)
GN
.DN
AB
.1 (B
1)
GN
.NA
B.1
(B1
)G
N.H
s_7
1_
TF.1
(B1
)G
N.A
_L
.08
(B1
)G
N.N
AB
.6 (B
1)
GN
.NA
B.6
(C9
)G
N.T
F.5 (C
12
)G
N.N
AB
.5 (D
3)
GN
.NA
B.2
(D4
)G
N.H
s_7
1_
TF.1
(D4
)G
N.b
ind
ing
.07
-2 (D
5)
GN
.NT
B.2
(D5
)G
N.D
NA
B.1
(D5
)G
N.N
AB
.1 (D
5)
GN
.A_
L.0
8 (D
5)
GN
.NA
B.6
(D5
)G
N.N
AB
.5 (D
6)
GN
.bin
din
g.0
7-2
(D9
)G
N.N
TB
.2 (D
9)
GN
.DN
AB
.1 (D
9)
GN
.NA
B.1
(D9
)G
N.A
_L
.08
(D9
)G
N.N
AB
.5 (D
9)
GN
.NA
B.6
(C9
)G
N.N
AB
.2 (E
8)
GN
.NA
B.2
(F1
)G
N.b
ind
ing
.07
-2 (F
12
)G
N.N
TB
.2 (F
12
)G
N.D
NA
B.1
(F1
2)
GN
.NA
B.1
(F1
2)
GN
.A_
L.0
8 (F
12
)G
N.N
AB
.5 (F
12
)G
N.N
AB
.6 (F
12
)G
N.b
ind
ing
.07
(G1
2)
GN
.NA
B.3
(G1
2)
GN
.NT
B.7
(G1
2)
GN
.TF.4
(G1
2)
GN
.NA
B.4
-1 (H
11)
GN
.NA
B.4
-2 (H
11)
GN
.NA
B.4
-3 (H
11)
GN
.bin
din
g.0
7-2
(C1
)G
N.N
AB
.2 (C
1)
GN
.DN
AB
.1 (C
1)
GN
. NA
B.1
(C1
)G
N.H
s_7
1_
TF.1
(C1
)G
N.A
_L
.08
(C1
) G
N.N
AB
.6 (C
1)
GN
. NA
B.2
(C1
2)
GN
.NA
B.6
(D9
)G
N.N
AB
.5 (E
3)
GN
. NA
B.2
(E4
)G
N.H
s_7
1_
TF.1
(E4
)G
N.b
ind
ing
.07
-2 (E
5)
GN
.NA
B.2
(E5
)G
N.D
NA
B.1
(E5
)G
N. N
AB
.1 (E
5)
GN
.A_
L.0
8 (E
5)
GN
.NA
B.6
(E5
)G
N.N
AB
.5 (E
6)
GN
.bin
din
g.0
7-2
(E9
)G
N.N
AB
.2 (E
9)
GN
.DN
AB
.1 (E
9)
GN
. NA
B.1
(E9
)G
N.A
_L
.08
(E9
)G
N.N
AB
.5 (E
9)
GN
. NA
B.2
(F8
) G
N. N
AB
.2 (G
1)
GN
.bin
din
g.0
7-2
GN
.NA
B.2
(G1
2)
GN
.DN
AB
.1 (G
12
)G
N. N
AB
.1 (G
12
)G
N.A
_L
.08
(G1
2)
GN
.NA
B.5
(G1
2)
GN
.NA
B.6
(G1
2)
Rela
tive
luc
activ
ity (%
)
Luc. Luc. + pSUPER -FL774
Luc. + pSUPER-FL774 + pSUPER-retro-puro
Luc. + pSUPER-FL774 + EIF2C2
Control.2 (position)
A1 A12 C4� C5� C9� E12� H9 F12 H10 B1 D5 D9 F12 H11 C1 E5 E9 G12 G22
! " # $ % & ' ( ) !* !! !"
+
,
-
.
/
0
1
2
.3435678
.34359:;<=
0
20
40
60
80
100
120
1000
10000
100000
のこり① Re
lativ
e lu
c ac
tivity
(%)
Renilla
luc
activ
ity Control
pSUPER-retro-puro EIF2C2
ABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHEFGH
1 2 3 4 5 6 7 8 9 10 11 12
061110
STK25 TAX1BP3
CIT
DMC1
ACTL6A
0
20
40
60
80
100
120
1000
10000
100000
のこり② Re
lativ
e lu
c ac
tivity
(%)
Renilla
luc
activ
ity Control
pSUPER-retro-puro EIF2C2
ABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHEFGH
1 2 3 4 5 6 7 8 9 10 11 12
061110
SFRS9�
AEBP1 AFF4
BAZ1B BHLHB2
D4S234E
DHX36
FLJ21839
0
20
40
60
80
100
120
1000
10000
100000
のこり③ Re
lativ
e lu
c ac
tivity
(%)
Renilla
luc
activ
ity Control
pSUPER-retro-puro EIF2C2
ABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHABCDEFGHEFGH
1 2 3 4 5 6 7 8 9 10 11 12
061110
INTS6
PAPD1
ZNF549
ZNF554
STK31