nmi tt pharmaservices proteinprofiling brochure ps/pdf protein profiling/n… · nmi tt...
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µ µ µ µ
µ
µ µ µ
0.0000001
0.000001
0.00001
0.0001
0.001
0.01
0.1
1
-5 -4 -3 -2 -1 0 1 2 3 4 5p-v
alu
e (
T-t
est
)
log2 fold-change to parental cell line
Lapatinib resistant lysates - DigiWest
Aurora A - phospho Thr288
Aurora A (AIK)
Aurora B (AIM1)
Caspase 7
Caspase 9
Caveolin-1
cdc2 (CDK1)
CDK6
Cyclin B1
Cytokeratin 8
DUSP6 (MKP3)
DUSP9 (MKP4)
EGFR
EGFR - phospho Tyr1068
EGFR - phospho Tyr845
EphA2
Erk1/2 (p44/42) - pT202/Y204
Her2
IRAK4 Phospho T345/S346
LATS2
MKK6
p21/Waf1/Cip1
p27(Kip1) (CDKN1B) - pS10
p27(Kip1) (CDKN1B)
p53
Pyk2 / FAK 2
RSK 1
RSK 1/2/3
Src - phospho Tyr416
STEP (PTPN5)
YAP - phospho Ser127
H29
2 P
are
nta
l Re
p 1
H29
2 P
are
nta
l Re
p 2
H29
2 P
are
nta
l Re
p 3
H29
2 R
esi
stan
t R
ep 1
H29
2 R
esi
stan
t R
ep 2
H29
2 R
esi
stan
t R
ep 3
0.000001
0.00001
0.0001
0.001
0.01
0.1
1
-10 -5 0 5
p v
alu
e (
T t
est
)
Log2 fold changes control vs. Lapatinib resistant
DigiWest KB Pulldown Analysis
EGFR
EphA2 pS897
Cyclin B1
EphA4
0.000001
0.00001
0.0001
0.001
0.01
0.1
1
-5 -4 -3 -2 -1 0 1 2 3
Log
(10
) p
-valu
e (
AN
OV
A)
Triplicate KB-PD (Contr and Lapatinib resistant) – Kinases
Lysate
Kinobeads 2. DigiWest
3. DigiWest1. MassSpec
H292 parental H292 resistant+ 1uM Lapa nib
-4
-3
-2
-1
0
1
2
3
4
EG
FR
PK
C
IKK
cdc2
(CD
K1
)
Au
rora
A (A
IK)
Ep
hA
2
LIM
K1
Au
rora
B (A
IM1
)
AS
K1
Ca
sein
kinase
1 e
psilo
n
Ep
hB
4
SIK
3
LR
RK
2
Ab
l2
IRA
K4
Pyk2
/ FA
K 2
LA
TS
2
RS
K 1
CD
K6
MK
K6
Ep
hB
3
Fyn
Ep
hB
6
Ep
hA
4L
og
2 f
old
ch
an
ge
s
KinoBead pulldown analysis: MS vs. DigiWest
DigiWest
Mass Spectrometry
0.0001
0.001
0.01
0.1
1
-5 -3 -1 1 3 5
p v
alu
e (
T t
est
)
Log2 fold changes control vs. Lapatinib resistant
DigiWest Lysate Analysis
DUSP6 (MKP3)
IRAK4 pT345/S346
Aurora A - pT288
DUSP9 (MKP4)
CDK6
Anderes Bild
P
P
PP
Phosphorylation
Predicted inhibition
Effect not predicted
Predicted activation
Downregulation
Upregulation
P
0.000001
0.00001
0.0001
0.001
0.01
0.1
1
-5 -4 -3 -2 -1 0 1 2 3Log
(10
) p
-va
lue
(A
NO
VA
)
Log2 fold change
Triplicate KB-PD (Contr and Lapatinib resistant) – Kinases
0.000001
0.00001
0.0001
0.001
0.01
0.1
1
-5 -4 -3 -2 -1 0 1 2 3Lo
g(1
0)
p-v
alu
e (
AN
OV
A)
Log2 fold change
Triplicate KB-PD (Contr and Lapatinib resistant) – Kinases
EGFR IKBKE
AURKA
EPHB3
EphA4Fyn
RPS6KA1
-4
-3
-2
-1
0
1
2
3
4
EG
FR
PK
C
IKK
cdc2
(CD
K1
)
Auro
ra A
(AIK
)
Eph
A2
LIM
K1
Auro
ra B
(AIM
1)
AS
K1
Ca
sein
kina
se 1
Eph
B4
SIK
3
LR
RK
2
Abl2
IRA
K4
Pyk2
/ FA
K 2
LA
TS
2
RS
K 1
CD
K6
MK
K6
Eph
B3
Fyn
Eph
B6
Eph
A4
Lo
g2 f
old
ch
an
ge
s
KinoBead pulldown analysis: MS vs. DigiWest
DigiWest
Mass Spectrometry
1E-08
0.0000001
0.000001
0.00001
0.0001
0.001
0.01
0.1
1
-10 -8 -6 -4 -2 0 2 4p-v
alu
e (
T-t
est
)
log2 fold-change to parental cell line
Lapatinib resistant KB pulldown - DigiWest
EGFR
EphA2 pS897
Cyclin B1Fyn
EphA2
IRAK4 pT345/S346
EGFR
DUSP6 (MKP3)
Aurora A pT288
DUSP9 (MKP4)
ERK1/2 pT202/Y204 p27(Kip1) pS10
CDK6
Aurora A - pT288
0.0000001
0.000001
0.00001
0.0001
0.001
0.01
0.1
1
-5 -4 -3 -2 -1 0 1 2 3p-v
alu
e (
T-t
est
)
log2 fold-change to parental cell line
Lapatinib resistant lysates - DigiWest
DUSP6 (MKP3)
DUSP9 (MKP4)
ERK1/2 pT202/Y204p27(Kip1) pS10
CDK6
p53
µ
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