0 4 8 12 24 48 (hr)
19
0 4 8 12 24 48 (hr) Cleaved Caspase-9 Caspase-8 100 nM Paclitaxel 0 2 4 6 8 C ontrol Paclitaxel 100 nM R FU (A rbitrary U nits) C aspase-9 activity C aspase-8 activity MCF-7 0 10 20 30 40 50 60 70 80 90 100 Cont rol Pacl i t axel Z-VAD- FM K + Pacl itaxe l Z-IETD -FM K + Pacl itaxel Z-LE HD -FM K + P a clitaxel % A nnexin V-positive cells Figure S1a
description
100 nM Paclitaxel. 0 4 8 12 24 48 (hr). Cleaved Caspase-9. Caspase-8. MCF-7. Figure S1a. **. *. MCF-7. Figure S1b. Paclitaxel 100 nM. - + - + - +. Bad siRNA. - - + + - -. Scrambled siRNA. - PowerPoint PPT Presentation
Transcript of 0 4 8 12 24 48 (hr)
0 4 8 12 24 48 (hr)
Cleaved Caspase-9
Caspase-8
100 nM Paclitaxel
0
2
4
6
8
Contro
l
Paclit
axel
100
nM
RF
U (
Arb
itra
ry U
nit
s)
Caspase-9 activity Caspase-8 activity
MCF-7
0
10
20
30
40
50
60
70
80
90
100
Contro
l
Paclit
axel
Z-VAD-F
MK +
Pac
litax
el
Z-IETD-F
MK +
Pac
litax
el
Z-LEHD-F
MK +
Pac
litax
el
% A
nn
exin
V-p
osi
tive
cel
ls
Figure S1a
MCF-7
Figure S1b
0
10
20
30
40
50
60
70
80
90
100
Contro
l
Paclit
axel
Bim s
iRNA
Paclit
axel +
Bim
siR
NA
Paclit
axel +
ABT-7
37+ B
im s
iRNA
Bmf s
iRNA
Paclit
axel +
Bm
f siR
NA
Paclit
axel +
ABT-7
37+ B
mf s
iRNA
ABT-737
ABT-737
+ Bim
siR
NA
ABT-737
+ Bm
f siR
NA
Paclit
axel +
ABT-7
37
% A
nn
exin
V-p
osi
tive
cel
ls
***
- - + + - -
- + - + - +Paclitaxel 100 nM
Bad siRNA
Scrambled siRNA - - - - + +
Bad
Actin
IP: Bcl-2
IB: Bcl-2
Input
IB: Bcl-2
Bcl-2
Bcl-2
Actin
IP: Bcl-2
IB: Bim
BimELBimLBimS
MCF-7
Figure S1c
0 4 8 12 24 48 (hr)
Cleaved Caspase-9
Caspase-8
100 nM Paclitaxel
Cleaved Caspase-3
0
2
4
6
8
RF
U (
Arb
itra
ry U
nit
s)
Caspase-9 activity Caspase-8 activity Caspase-3 activity
T47D
0
10
20
30
40
50
60
70
80
90
100
Contro
l
Paclit
axel
Z-VAD-F
MK +
Pac
litax
el
Z-IETD-F
MK +
Pac
litax
el
Z-LEHD-F
MK +
Pac
litax
el
% A
nn
exin
V-p
osi
tive
cel
ls
Figure S1d
0 4 8 12 24 48 (hr)
Cleaved Caspase-9
Caspase-8
100 nM Paclitaxel
Cleaved Caspase-3
0
2
4
6
8
RF
U (
Arb
itra
ry U
nit
s)
Caspase-9 activity Caspase-8 activity Caspase-3 activity
MDA-MB-468
0
10
20
30
40
50
60
70
80
90
100
Contro
l
Paclit
axel
Z-VAD-F
MK +
Pac
litax
el
Z-IETD-F
MK +
Pac
litax
el
Z-LEHD-F
MK +
Pac
litax
el
% A
nn
exin
V-p
osi
tive
cel
ls
Figure S1e
0 4 8 12 24 48 (hr)
Cleaved Caspase-9
Caspase-8
100 nM Paclitaxel
Cleaved Caspase-3
0
2
4
6
8
RF
U (
Arb
itra
ry U
nit
s)
Caspase-9 activity Caspase-8 activity Caspase-3 activity
BT20
0
10
20
30
40
50
60
70
80
90
100
Contro
l
Paclit
axel
Z-VAD-F
MK +
Pac
litax
el
Z-IETD-F
MK +
Pac
litax
el
Z-LEHD-F
MK +
Pac
litax
el
% A
nn
exin
V-p
osi
tive
cel
ls
Figure S1f
IP: Bcl-xL
IB: Bcl-xLBcl-xL
Bim EL
Bim L
Bim S
IP: Bcl-xL
IB: Bim
Mcl-1
Paclitaxel 100 nM
0 4 8 12 [h]
Bim EL
Bim L
Bim S
IP: Mcl-1
IB: Bim
Paclitaxel 100 nM
0 4 8 12 [h]
IP: Mcl-1
IB: Mcl-1
Figure S2a
MCF-7
Figure S2b
Pre
-IP
Po
st-I
P
BimELBimLBimS
HM LM S-100
Untreated Taxol 12 h
CoxIV
BiP/Grp78
BimEL
BimL BimS
1 0.43
Actin
Ratio
Paclitaxel 100 nM
- +
Pre
-IP
Po
st-I
P
Actin
BimELBimLBimS
Untreated
1 1.39Ratio
HM LM S-100
MCF-7 MCF-7MCF-7
- + - +
Paclitaxel 100 nM
MCF-7 MCF-7 Bcl-2+
FLAG-Bcl-2
Bcl-2FLAG-Bcl-2
BimEL
BimL BimS
IP: FLAG
IB: Bcl-2
Input
IB: Bcl-2
IP: FLAG
IB: Bim
Bmf
Bmf
IP: FLAG
IB: Bmf
Input
IB: Bmf
BimELBimL
BimS
Input
IB: Bim
Actin
IP: FLAG
IB: Bax
IP: FLAG
IB: Puma
*
IP: FLAG
IB: Bak
IP: FLAG
IB: Bad
- + - +
Paclitaxel 100 nM
MCF-7 MCF-7 Bcl-2+
Bad
Puma
Bax
Bak
Figure S2c
Figure S2d
IP: Bcl-2
IB: BaxBax
IP: Bcl-2
IB: Bak
Paclitaxel 100 nM
- +
IP: Bcl-xL
IB: Bax
IP: Bcl-xL
IB: Bak
Paclitaxel 100 nM
- +
IP: Mcl-1
IB: Bax
IP: Mcl-1
IB: Bak
Paclitaxel 100 nM
- +
Bak
Bax
Bak
Bax
Bak
MCF-7
0 4 8 12 24 48 0 4 8 12 24 48 (hr)
MitoCyto
Cytochrome c
CoxIV
T47D
Figure S3a
0 4 8 12 24 48 0 4 8 12 24 48 (hr)
MitoCyto
MDA-MB-468
Figure S3b
Cytochrome c
CoxIV
0 4 8 12 24 48 0 4 8 12 24 48 (hr)
MitoCyto
BT20
Figure S3c
Cytochrome c
CoxIV
0 4 8 12 24 48 (hr)
Bax
Bak
IP: Bax 6a7
IB: Bax N20
IP: Bak Ab-2
IB: Bak G23
100 nM Paclitaxel
T47D
Input
IB: Bax N20
Input
IB: Bak G23
Bax
Bak
Actin
Figure S4a
0 4 8 12 24 48 (hr)
Bax
Bak
IP: Bax 6a7
IB: Bax N20
IP: Bak Ab-2
IB: Bak G23
100 nM Paclitaxel
MDA-MB-468
Input
IB: Bax N20
Input
IB: Bak G23
Bax
Bak
Actin
Figure S4b
0 4 8 12 24 48 (hr)
Bax
Bak
IP: Bax 6a7
IB: Bax N20
IP: Bak Ab-2
IB: Bak G23
100 nM Paclitaxel
BT20
Input
IB: Bax N20
Input
IB: Bak G23
Bax
Bak
Actin
Figure S4c
Puma
Actin
- - + + - -
- + - + - +Paclitaxel 100 nM
Puma siRNA
Scrambled siRNA - - - - + +
MDA-MB-468
0
10
20
30
40
50
60
70
80
90
100
Contro
l
Paclit
axel
Puma
siRNA
Paclit
axel +
Pum
a siR
NA
Scram
bled
siRNA
Paclit
axel +
Scr
amble
d siR
NA
% A
nn
exin
V-p
osi
tive
cel
ls
Figure S5a
Puma
Actin
- - + + - -
- + - + - +Paclitaxel 100 nM
Puma siRNA
Scrambled siRNA - - - - + +
BT20
0
10
20
30
40
50
60
70
80
90
100
Contro
l
Paclit
axel
Puma
siRNA
Paclit
axel +
Pum
a siR
NA
Scram
bled
siRNA
Paclit
axel +
Scr
amble
d siR
NA
% A
nn
exin
V-p
osi
tive
cel
ls
Figure S5b
Puma
Actin
- - + + - -
- + - + - +Paclitaxel 100 nM
Puma siRNA
Scrambled siRNA - - - - + +
MCF-7
0
10
20
30
40
50
60
70
80
90
100
Contro
l
Paclit
axel
Puma
siRNA
Paclit
axel +
Pum
a siR
NA
Scram
bled
siRNA
Paclit
axel +
Scr
amble
d siR
NA
% A
nn
exin
V-p
osi
tive
cel
ls
Figure S5c
