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Copyright 2016 The Author(s). 

This is an Accepted Manuscript; not the final Version of Record. Archiving permitted only in line with the 

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Cite as Biochemical Journal (2016) DOI: 10.1042/BCJ20160137 

BIOCHEMICAL JOURNAL

ACCEPTED MANUSCRIPT 

10.1042/BCJ20160137

. Please cite using the DOI 

10.1042/BCJ20160137

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in vivo

in vitro

P

P

P

P

P

P

P

P

P

P

Figure 1.

A

μM



0

0.0005


0.001

0.0015


0.002

0.0025


0.003

0.0035


0.004

0.0045


0.005

0

50



100

Insulin secre

on

 p

er total 



p

rotein 


(n

g/

μ



g)

H

2



O

2

Control siRNA



SCGN siRNA

B

*



**

*

0



0.0005

0.001


0.0015

0.002


0.0025

0.003


0.0035

0

2.8



16.8

Insulin secre

on

 p

er total 



p

rotein 


(n

g/

μ



g)

Glucose


Control siRNA

SCGN siRNA

***

***


***

mM

tubulin



CTL

SCGN


siRNA

An - -


An -SCGN

CTL


SCGN

siRNA


An - -tubulin

An -SCGN


CTL

SCGN


siRNA

An - -tubulin

An -SCGN

C

0



1

2

3



4

5

6



7

8

9



10

3.3


16.8

In

su

lin

 r

e

le

ase

 p

e

r isle

ts

 (n

g/

isle

t)

Control siRNA

SCGN siRNA

*

Glucose



mM

B

0

16.8



CTL

SCGN


CTL

SCGN 


 An - -tubulin

 An -SCGN

5.4

30

KCl



siRNA

CTL


SCGN

CTL


SCGN 

Glucose


mM

A

0



0.0005

0.001


0.0015

0.002


0.0025

0.003


0.0035

5.4


30

Insulin secre

on

 p

er total 



p

rotein 


(n

g/

g)



Control siRNA

SCGN siRNA

KCl

mM

0



0.0002

0.0004


0.0006

0.0008


0.001

0.0012


0

16.8


Insulin secre

on

 p



er total 

p

rotein 



(n

g/

g)



Control siRNA

SCGN siRNA

Glucose

mM

*



Figure 2.

0

0.0005



0.001

0.0015


0.002

0.0025


0.003

0

10



20

30

40



Ac

cu

m

u

late

d

 in

su

lin

 se

cr

e

on per tota

l protei



(ng/

g)

Time, min

Control siRNA

SCGN siRNA

***


*

0

SCGN siRNA



16.8

Control siRNA

Glucose

(mM)


C

CTL


SCGN

An - -tubulin

An -SCGN

siRNA


N

N

N



PM           IgG SCGN       

SCGN


IgG

SCGN     IgG SCGN

Input         Output

IP

IgG SCGN 



A

C

-ac n



IP

58

46



30

25

- -ac n



-SCGN

- -ac n


-SCGN

B

Figure 3.



AVFPSIVGRPR 599.8804(2+)

mass


-0

50

100



150

200


250

300


350

400


450

500


550

600


650

700


750

800


850

900


950

1000


1050

1100


1150

%

0



100

5.86e3


881.5911

120.0866


70.0696

60.0574


72.0852

864.5797


567.3710

143.1257


272.1878

185.1036


245.1703

212.1519


468.2967

441.2512


280.1889

428.2893


314.1694

369.2815


397.2852

485.3235


524.3616

725.4980


584.3970

680.4709


628.4256

784.5338


821.5545

1198.7672

1028.6678

882.5065


1011.6394

927.6158


1029.5737

1042.6803

1043.5668

1167.6589

(P60710) Ac n, cytoplasmic 1 (Beta ac n) 

ACTB_Mouse Mass: 41710 Score: 99 Queries matched: 3

An body

An body


Immunopre

cipitated 

IP

IgG


SCGN     IgG

SCGN


Glucose          0                16.8        mM

- -ac n


-SCGN

A

B



F-ac n

Merge


SCGN

100


0

Ampli ed


- -ac n

-SCGN


H

2

O



(μM)


C

Figure 4.

0

5

10



15

20

25



30

35

40



1

2

3



4

5

6



7

8

Rela



ve in

te

n

sit



ra

o of

 S

C

GN (%)

Frac on no.

DMSO


Ionomycin

0

5



10

15

20



25

30

35



40

45

1



2

3

4



5

6

7



8

Rela

ve in

te

n

sit



ra

o of

 ac

n (%)

Frac on no.

DMSO


Ionomycin

An -SCGN


An - -ac n

DMSO


Ionomycin

1    2    3    4    5    6    7    8

Top        frac on no.      Bo om

1    2    3    4    5    6    7    8

Top        frac on no.      Bo om

DMSO


Ionomycin

IgG


SCGN     IgG

SCGN


Input

Glucose            0               16.8                  0                16.8   mM

An body

IgG


SCGN     IgG

SCGN


Output

An body


80

90

100



110

120


130

140


150

160


170

180


0

16.8


-a

c

n/SC



G

N

 ra



o (%

)

mM



Glucose

*


Control                

SCGN


An - -ac n

G-ac n   F

-

ac n


siRNA

G-ac n   F-ac n

An - -ac n

An -SCGN


Total ac n

Control  SCGN

A

B

0



0.2

0.4


0.6

0.8


1

1.2


F-

ac

n to 



G

-ac


n ra

o (a


rbi

tra


ry

 uni


ts

)

Control siRNA



SCGN siRNA

*

DAPI



siRNA

F-ac n


Merge

SCGN


Ampli ed

Control


0

100 Control

SCGN

0

100



SCGN

siRNA


Figure 5.

H

2



O

(μM)



Control siRNA

SCGN siRNA

A

C

DAPI



siRNA

Merge


SCGN

Control


0

100 Control

SCGN

0

100 SCGN



Paxillin

An -E-Cadherin

An -N-Cadherin

siRNA CTL   SCGN

B

An -SCGN


An -tubulin

Figure 6.

H

2

O



(μM)


Figure 7.

An -p-ERK

An -ERK

An -p-AKT



An -AKT

An -SCGN


An -tubulin

An -p-paxillin

An -paxillin

0              5            15           30

min

0             5            15          30



H

2

O



2

An -p-FAK

An -FAK

Control


SCGN

siRNA


0             2.8         16.8          30

Glucose


mM

0            2.8         16.8          30

An -SCGN

An -p-FAK

An -FAK

An -p-paxillin



An -paxillin

An -p-ERK

An -ERK

An -p-AKT



An -AKT

Control


SCGN

siRNA


0             2.8         16.8          30

Glucose


mM

0            2.8         16.8          30

0

50

100



150

200


250

300


0

2.8 16.8 30



Ph

o

sp

h

o

-:To

tal FAK 

(%

)

Control siRNA

SCGN siRNA

0

100



200

300


400

500


600

0

2.8 16.8 30



P

h

os

pho-

:T

otal

 P

axi

lli

n

 (%)

Control siRNA

SCGN siRNA

0

50



100

150


200

250


300

0

2.8 16.8 30



Ph

o

sp

h

o

-:To

tal Akt (%

)

Control siRNA

SCGN siRNA

0

50



100

150


200

250


300

0

2.8 16.8 30



Phospho-:Total

 E

R



(%)

Control siRNA

SCGN siRNA

*

*



**

**

* *



*

***


******

Control


0             20            50          100

H

2



O

2

M



SCGN

An -SCGN


0             20            50          100

An -p-FAK

An -FAK

siRNA


An -p-paxillin

An -paxillin

An -p-ERK

An -ERK


An -p-AKT

An -AKT


Control

0             20            50          100

H

2

O



2

M

SCGN



0             20            50           100

siRNA


0

50

100



150

200


250

300


350

400


0

20 50 100



Ph

o

sp

h

o

-:To

tal FAK 

(%

)

Control siRNA

SCGN siRNA

0

50



100

150


200

250


0

20 50 100



Ph

o

sp

h

o

-:To

tal Paxillin

 (%

)

Control siRNA

SCGN siRNA

0

100



200

300


400

500


600

0

20 50 100



Ph

o

sp

h

o

-:To

tal Akt (%

)

Control siRNA

SCGN siRNA

0

200



400

600


800

1000


1200

1400


1600

1800


0

20 50 100



P

h

os

pho-

:T

o

ta

l E

R

K

 (%)

Control siRNA

SCGN siRNA

**

*



*

*

*



*

*

***



***

C

A



B

Control


SCGN

siRNA


Glucose

Glucose


Glucose

Glucose


mM

mM

mM



mM

H

2



O

2

H



2

O

2



H

2

O



2

H

2



O

2

M



M

M

M



Control               SCGN

DMSO


Glucose

Latrunculin B

-

+          -



+

-

+          -



+

Control                SCGN

An -p-ERK

An -ERK


An -p-FAK

An -FAK


An -SCGN

siRNA


Figure 8.


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