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FSH
FSHR
LH
LHR
Gs
Gi
Gq
Gh
EGFR
Src
Transactivation
PLCδ
IP3
cAMP
PKAERK EPAC PI3K
Aktp38
Cdk4
Appl1
ßarr
Rps6
PDE
Figure 1
k0
k1
Ordinary differential equation (ODE)
Mass action law:
k0
k1Mass action law:
(1)
(2)
(3)
A
B
A B
A B
C
k0
k1
k2 k3
k4
C DA B
C D E
F G
[B]
Time
Simulation
δ1
δ2
δ3
δ4δ5
Experimentalpoints
E Initial parameter values (k0, ...,kn)
Simulation
Compute errorChange (k0,..., kn)
Done !
Error=0 ?
δ0B
B
B
B
BB
error =f(δ0, δ1, δ2, δ3, δ4, δ5 )B B B B B B B
pER
K (
µm
ol/L
)
[Ang]
Figure 3
k0k1k2k6
k3
k7
k4
k8
k10
k11 k13 k12 k14 k20 k24 k19 k23
k17
k18
k5
k9
k22k21
k25
k16
k15
Model equations Conservation laws
Biological conditions• Condition 1: control• Condition 2: PKC inhibition, k29 k33• Condition 3: siRNA Barr2, k31 k34• Condition 4: siRNA GRK23, k10 k35• Condition 5: siRNA GRK56, k18 k36
A
B
C
[C]
Time
Stimulation
A B
Time
Stimulation
[C]
Figure 4
Epac
RFP GFPEpac
RFP GFP
BRET
cAMP
BRET
A
BRETBRET
Rluc
Gα
Ven
Gß/γ
Rluc
Gα
Ven
Gß/γ
B
BRET
cha
nge
Time (min)
FSH
Time (min)
LH
Time (min) Time (min)
GFP
BRET
C
AEQ
Ca2+
GFPAEQ
D
BRETßarryPet
Rluc Rlucßarr
yPet
+hFSH +hCGTime (s) Time (s)
Time (min) Time (min)
+hFSH +hCG
Figure 5
% o
f res
pons
e
log([hFSH]) log([hCG])
Figure 6
Effector
Kd
τ
Ligand
[Ligand]
[Effector]
Emax
EC50
A
Kd
Ligand
B
Effector1 Effector2
[Ligand]
[Effector]
Emax-1
EC50
-1
Emax-2
EC50
-2
Kd
Ligand
C
Effector1 I1
Effector2
τ1 τ2
Kd
FSH
D
Gαs ß-arrestin
τ(G,FSH) τ(ß,FSH)
Kd
L1
Gαs ß-arrestin
τ(G,L1) τ(ß,L1)
τ(ß,L1)τ(G,L1)
τ(ß,FSH)τ(G,FSH)>
Kd
L2
Gαs ß-arrestin
τ(G,L1) τ(ß,L1)
τ(ß,L2)τ(G,L2)
τ(ß,FSH)τ(G,FSH)<
Figure 7
Time (min)
activ
e G
(µm
ol/L
)
AT1-
ß-ar
rest
in 2
sig
nalin
g co
mpl
ex (µ
mol
/L)
Time (min)
Figure 8