Co

ntr

ol

siR

NA

BR

G1

siR

NA

PTEN –

Akt –

p-Akt –

p-GSK-3–

Cyclin D1 –

BRG1 –

-actin –

SW480

Supplemental data S1

Supplemental data S1. Effects of transduction of BRG1 siRNA into CRC-derived SW480 cells. Knockdown of BRG1 was confirmed by Western blot. Altered expressions of the PI3K-Akt signaling -related molecules (PTEN, Akt, p-Akt, p-GSK-3 and cyclin D1) were investigated. -actin was used as a loading control.

Supplemental data S2

Watanabe et al.

PIK

3CB

rel

ativ

e ex

pre

ssio

n(/

Cyc

lop

hil

in)

Control siRNA BRG1 siRNA

PIK

3CG

rel

ativ

e ex

pre

ssio

n(/

Cyc

lop

hil

in)

Control siRNA BRG1 siRNA

PIK

3CA

rel

ativ

e ex

pre

ssio

n(/

Cyc

lop

hil

in)

Control siRNA BRG1 siRNAP

IK3C

D r

elat

ive

exp

ress

ion

(/C

yclo

ph

ilin

)

Control siRNA BRG1 siRNA

PD

K1

rela

tive

exp

ress

ion

(/C

yclo

ph

ilin

)

Control siRNA BRG1 siRNA

*

PIK

3D1

rela

tive

exp

ress

ion

(/C

yclo

ph

ilin

)

Control siRNA BRG1 siRNA

*

Supplemental data S2. Results of qRT-PCR analysis. Altered expressions of the PI3K-Akt signaling-related genes (PDK1, PIK3CA, PIK3CB, PIK3CD, PIK3CG and PIK3D1) were examined. The cyclophilin mRNA levels were used as a control.

Supplemental data S3

Watanabe et al.

A

1. Control siRNA

2. BRG1 siRNA

1

-catenin –

-actin –

2

Cytosol

1 2

Membrane/organelle

1 2

Nucleus

1 2

Cytoskeleton/matrix

B

Cyc

lin

D1

mR

NA

lev

el(/

Cyc

lop

hil

in)

Control siRNA BRG1 siRNA0

0.5

1

1.5

2

Supplemental data S3. (A) The amount of -catenin in subcellular fractions in the BRG1 siRNA- and control siRNA-transfected DLD-1 cells. (B) The cyclin D1 mRNA levels in the BRG1 siRNA- and control siRNA-transfected DLD-1 cells.