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% of lethality of animals

Cucurbitacin B (Cuc B)

Cucurbitacin E (Cuc E)

Fig. 1

20

40

60

80

100

EtOH

(80)

Ecd

(40)

CucE

(40)

CucB

(70)

CucB (0.1 mM)

(40)

CucB + Ecd

(70)

CLR

(65)

CucB + CLR

(50)

7dC

(40)

CucB + 7dC

(40)

L1

L1/L2

L2

L2/L3

L3

Prepupa

Pupa

Adult

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100

Control

80

% of animals

L1

60

L2

40

L3

Pupa

20

Lost

48h

72h

96h

120h

144h

168h

(50)

(78)

(90)

(72)

(80)

(20)

100

+ Cuc B

80

% of animals

L1

60

L2

L3

40

Pupa

20

Lost

48h

72h

96h

120h

144h

168h

(50)

(62)

(90)

(80)

(80)

(30)

Fig. 2

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Fig. 3

Control

**

Larval area (mm2)

0.1

1.0

Absorbance at 630 nm

0.2

CucB-fed

0.5

Control

CucB

Control

CucB

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% of lethality of animals

10

20

30

40

50

60

70

80

CLR

(30)

CucB

(50)

CucB+Ecd

(40)

Ecd

(30)

Pupa

Larval weight (mg)

(30)

Incomplete pupa

Fig. 4

90 100

EtOH

L3

1.5

1.0

b,c

0.5

0.0

48h

48h

Hours after L2/L3 molt

Adult

CucB

EtOH

Percentage of pupariation

100

80

EtOH (30)

60

CucB (25)

40

CucB+Ecd (25)

20

Ecd (29)

24h

48h

72h

96h

Hours after L2/L3 molt

120h

144h

ns

60

20E titer (pg / mg prepupa)

72h

40

20

EtOH

CucB

96h

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Fig. 5

ns

15

ns

Relative amount

ns

10

Control

CucB

25 mg / ml

(45 mM)

Control

CucB

2.5 mg / ml

(4.5 mM)

Control

CucE

25 mg / ml

(45 mM)

Control

CucE

2.5 mg / ml

(4.5 mM)

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Fig. 6

100

Relative activity (%)

80

60

CucB

CucE

40

17b-Estradiol

20

0.03

0.1

0.3

Concentration (mM)

10

30

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Drosophila melanogaster

Bombyx mori

Inhibition

Inhibition

Prothoracic glands

In vitro culture

Cucurbitacin B

Ecdysone biosynthesis

Larval development

No effect

Cucurbitacin E

...