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C B A DE F Fig. 1

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(1)

0 1 2 3

Pr o ge stero n e (fo ld s)

0 0.5 1 1.5

Es tr ad io l ( fo ld s)

FSH

(10 ng/ml)

ORX

(100 nM)

- -

+ + +

- -

+ MM Hctr1 Hctr2 Rpl19

422bp 350bp 195bp

Fig. 1 A

a a

b c

a a

b b

B

0 0.5 1 1.5 2

Pr o ge stero n e ( fo ld s)

a b

c

FSH

(10 ng/ml)

ORX

(100 nM)

Nog

(30 ng/ml)

DOR

(300 nM)

LDN

(300 nM) - - - - -

+ + + - - + + - - - + - - - -

+ + - + -

+ + - - +

b b

ab

C

0 0.1 0.2 0.3 0.4 0.5

cAMP le ve l ( nM )

FSH

(10 ng/ml)

ORX

(100 nM)

- -

+ + +

- - +

a a

b

D

c

E

mRN A le ve ls (/ Rp l19 )

0 100 200 300 400 500 0

1 2 3

0 5 10 15 20 25

0 1 2 3

Cyp11a

mR N A le ve ls ( / Rp l1 9 )

a

ab b

a

ab b

a

b

a

b b a

FSH ORX

+ + -

-

+ - - +

a

a

a

FSH ORX

+ + -

-

+ - - +

FSH ORX

+ + -

-

+ - - +

FSH ORX

+ + -

-

+ - - + 0 0.5 1 1.5 2

- +

ORX

(100 nM)

Fshr

mR N A le ve ls ( / Rp l1 9 )

F

Star

Hsd3b

c

Cyp19

(2)

0 0.5 1 1.5 2

- -

+ + - + +

-

a a

b a

ORX BMP-7

pSmad/tSmad1

IB: pSmad 1/5/9 IB: tSmad1

A

mRNA levels (/RPL19)

ORX

(100 nM)

BMP-6

(100 ng/ml)

SB408124

(10 μM)

- - -

+ - -

- - +

- + -

+ + -

+ + + 0

1 2 3 4

b

a

b

Id1

a a

a

mRNA levels (/RPL19)

- - -

- - + +

- - ORX

BMP-7 SB408124

- + -

+ + -

+ + + 0

1 2 3 4 5

b

a

ab

Id1

a a

b

B

pSmad/tSmad1

ORX

(100 nM)

BMP-6

(100 ng/ml)

- -

+ + - + +

-

a a

b a IB: pSmad

1/5/9 IB: tSmad1

0 0.5 1 1.5 2

Fig. 2

(3)

Fig. 3

0 0.5 1 1.5 0

0.5 1 1.5

0 0.5 1

1.5 Bmpr1a Bmpr1b

0 0.5 1 1.5

- + - +

- + Acvr1

mRNA levels (/Rpl19)

ORX

(100 nM)

** *

*

0 0.5 1

1.5

Acvr2a Bmpr2

0 0.5 1 1.5

- + - +

* *

0 0.5 1 1.5

2

Smad6 Smad7

mRNA levels (/Rpl19)

*

- +

+ BMP-6

(100 ng/ml)

+ BMP-7

(100 ng/ml)

mRNA levels (/Rpl19)

0 1 2 3

- +

**

0 0.5 1 1.5 2

- +

*

0 1 2 3

- +

*

0 0.5 1 1.5

0 0.5 1 1.5

0 0.5 1 1.5

- + - +

- +

BMP-6

(100 ng/ml)

- +

mRNA levels (/Rpl19)

** ** ** **

mRNA levels (/Rpl19)

BMP-7

(100 ng/ml)

ORX

(100 nM)

ORX

(100 nM)

ORX

(100 nM)

Hctr1 Hctr2 Hctr1 Hctr2

A

B

C

Smad6 Smad7

(4)

FSH

Progesterone induction

Orexin A BMPs

Id-1

cAM P

-

Granulosa cells

Smad

1/5/9

P OX1/OX2

FSHR

Steroidogenetic enzymes

StAR, P450scc, 3βHSD BMPRs

AC Nog

Smad

6/7

type-I type-II

DOR LDN SB

- -

+

Fig. 4

図

Fig. 3 0 0.511.500.511.500.511.5Bmpr1aBmpr1b 00.511.5-+-+-+Acvr1mRNA levels (/Rpl19)ORX(100 nM)****00.511.5 Acvr2a Bmpr200.511.5-+-+**00.511.52Smad6Smad7 mRNA levels (/Rpl19)*-++ BMP-6(100 ng/ml)+ BMP-7(100 ng/ml)mRNA levels (/Rpl19)0123-+**00.511.52-+*01

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