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