Tfa

m 1

g

empt

y v

ecto

r 1

g

Tfb

2m 1

g

Rel

ativ

e m

RN

A le

vel

0

100

200

50

150

**

p<0.05 vs empty vector*

Transfection

Tfa

m m

RN

A

Tfb

2m m

RN

A

Tfa

m a

nd

Tfb

2m m

RN

A

Tfa

m s

iRN

A #

1

Scr

ambl

e si

RN

A

Tfb

2m s

iRN

A #

10

0.75

0.5

0.25

1

* *

Tfa

m s

iRN

A #

2

Tfb

2m s

iRN

A #

2

* *

p<0.05 vs scramble siRNA*

Tfa

m m

RN

A

Tfb

2m m

RN

A

Transfection

Tfa

m a

nd

Tfb

2m m

RN

A

Tfam

Tfb2m

28S

Non

-tre

ated

Tfa

m s

iRN

A #

1

Scr

ambl

e si

RN

A

Tfb

2m

siR

NA

#1

Tfa

m s

iRN

A #

2

Tfb

2m

siR

NA

#2

Rel

ativ

e m

RN

A le

vel

Supplemental Figure 2

BC

A

Transfection

A

Supplemental Figure 3

Tra

nsla

tiona

l diff

usio

n tim

e (

s)

0

200

400

600

800

1000

1200

1400

-1769 -1739 -1709-1679-1649 -1619-1589 -1559 -1529-1499-1469 -1439 -1409

p<0.05

*

-1799-1829

RecombinantTFAM protein

(-) (+)

-1379

*

B

Tra

nsla

tiona

l diff

usio

n tim

e (

s)

0

200

400

600

800

1000

1200

1400

-1769 -1739 -1709-1679-1649 -1619-1589 -1559 -1529-1499-1469 -1439 -1409

*p<0.05

*

-1799-1829

RecombinantTFB2M protein

(-) (+)

-1379

*

1600

1800

*

*

Supplemental Figure 4

A B

Tfa

m s

iRN

A #

1

Scr

ambl

e si

RN

A 0

15

10

5

20

*

Tfb

2m s

iRN

A #

1

*

p<0.05 vs. scramble siRNA*

AT

P c

onte

nt

(nmol/mg tissue)

0

30

20

10

40

* *

p<0.05 vs. empty vector*

AT

P c

onte

nt(nmol/mg tissue)

Tfa

m 1

g

Em

pty

vect

or 1

g

Tfb

2m 1

g

Supplemental Figure 5

A B

Rel

ativ

e m

itoch

ondr

ial

AT

Pas

e 6/

8 m

RN

A le

vel

r = 0.49p < 0.01

0.25

0.5

0.75

1.0

1.25

2.0 2.51.51.00.5

Relative Tfam mRNA level

1.5

1.75

2.0

Rel

ativ

e m

itoch

ondr

ial

AT

Pas

e 6/

8 m

RN

A le

vel

r = 0.57p < 0.001

0.25

0.5

0.75

1.0

1.25

2.01.51.00.5

Relative Tfb2m mRNA level

1.5

1.75

2.0

D

Rel

ativ

e cy

toch

rom

e-b

mR

NA

leve

l

r = 0.48p < 0.01

0.25

0.5

0.75

1.0

1.25

2.0 2.51.51.00.5

Relative Tfam mRNA level

1.5

C

Rel

ativ

e cy

toch

rom

e-b

mR

NA

leve

l

r = 0.58p < 0.001

0.25

0.5

0.75

1.0

1.25

2.01.51.00.5

Relative Tfb2m mRNA level

1.5

Tra

nsla

tiona

l diff

usio

n tim

e (

s)

-170~-141

-164~-135

-158~-129

-152~-123

-146~-117

-140~-111

-134~-105

-131~-102

-128~-99

-125~-96

-122~-93

(-)(+)

p<0.05

0

500

1000

1500

2000

2500

3000*

***

*

Recombinant SP1 protein

A

C D

Tfa

m 0

.5

g

+S

p1

0.5

g

1

2

0.5

1.5

2.5

0

Em

pty

ve

cto

r 1

g

Tfa

m 1

g

Sp

1 1

g

3

Ser

ca2

gen

e tr

ansc

riptio

n(r

ela

tive

luci

fera

se u

nit)

* *

*p<0.05 vs. empty vector

0

200

400

600

800

1000

1200

TFAM only

TFAM+SP1

Tra

nsl

atio

na

l diff

usi

on

tim

e (s

) NS

Supplemental Figure 6

B

Ser

ca2

gen

e tr

ansc

riptio

n(r

ela

tive

luci

fera

se u

nit) p<0.05 vs. empty

vector in each group

* *

*

Serca2 Luc wild Serca2 Luc mutation -122

Serca2 Lucmutation -119

Em

pty

Tfa

m

Tfb

2m

Sp

1

Em

pty

Tfa

m

Tfb

2m

Sp

1

Em

pty

Tfa

m

Tfb

2m

Sp

1

Em

pty

Tfa

m

Tfb

2m

Sp

1Serca2 Luc

mutation -116

*

**

0

1

2

3

4

*

*

*

*

Tfam

Tfb2m

Bnp

Serca2a

ET

-1 1

00 n

mol

/L

NE

30

mol

/L

NE

100

m

ol/L

Veh

icle

Veh

icle

28S

Supplemental Figure 7

ET

-1 1

0 nm

ol/L

Veh

icle

NE

5

mol

/L

Veh

icle