WT control A3R KO control WT-TAC A3R KO-TAC

Fib

rosi

s C

ell

size

Figure S1

Figure S1. Representative staining for cell size and myocardial fibrosis in A3R KO mice and wild type (WT) mice under control conditions and after moderate TAC for 5 weeks, indicating less myocyte hypertrophy and myocardial fibrosis in A3R KO mice after moderated TAC as compared with WT mice.

Days after moderate TAC0 3 6 9 12 15

Per

cent

Sur

vivi

ng

30

60

90

120

ShamWt TAC (2/13)A3R KO + TAC (2/14)

Figure S2. The TAC induced mortality was not different between A3R KO and Wild type mice (Wt) mice.

Sham TAC-2dB

od

y w

eig

ht

(g)

5

10

15

20

25

Sham TAC-2d

Ve

ntr

icu

lar

mas

s (

mg

)

50

100

150 WTA1R KO

**

Sham TAC-2dRa

tio

of

Ve

ntr

icu

lar

ma

ss

to b

od

y w

eig

ht

(mg

/g)

2

4

6

* *n=11

n=10

n=15

n=18

n=11

n=10

n=15

n=18

n=11

n=10

n=15

n=18

A B C

Figure S3. A1R KO had no effect on the increase of ventricular mass (A) or the ratio of ventricular mass to body weight 2 days after severe TAC. *P<0.05 compared to the corresponding control.

Days after TAC 0 10 20 30

Per

cen

t S

urv

ivin

g30

60

90

120

Sham (n=10)WT TAC (n=16/28)A1R KO TAC (10/32)

p<

0.05

Figure S4. A1R KO significantly increased mortality in mice subjected to severe TAC for 4 weeks. A: severe TAC-induced survival rate in WT and A1R KO mice.

Hea

rt r

ate

(b

eats

/min

)

ab c

Su

rger

y

Hours

Survived

a

b

cS

urg

ery

died

Hea

rt r

ate

(b

eats

/min

)

Hours

Figure S5: ECG telemetry shows progressive bradycardia in WT and A1R KO mice (C) after severe TAC.

WT control CD73 KO control WT TAC-4W CD73 KO TAC-4WA

Control TAC-4W

LV

eje

ctio

n f

ract

ion

(%

)

15

30

45

60

75

90

*

Control TAC-4W

LV

dia

met

er a

t en

d s

yst

ole

(m

m)

2

4

6

***

Control TAC-4W

LV

dia

met

er a

t en

d d

iast

ole

(m

m)

2

4

6

*##

#

Control TAC-4WHea

rt r

ate

(b

eats

/min

)

200

400

600

800 WTCD73 KO

**n

=8

n=

8

n=

11

n=

11

n=

8

n=

8

n=

11

n=

11

n=

8

n=

8

n=

11

n=

11

n=

8

n=

8

n=

11

n=

11

B C D E

Figure S6. CD73 KO significantly exacerbated the decrease of LV ejection fraction (A), increase of LV end systolic diameter (B), and increase of LV end diastolic diameter (C) produced by 4 weeks of moderate TAC. CD73 KO had no significant effect on heart rate under either control conditions or after moderate TAC (D).

TNF

β-MHC

ANF

A

Wild typeControl

Wild type TAC-4W

CD73 KOControl

CD73 KOTAC - 4W

3-NT

Control TAC-4W

Myo

card

ial A

NP

0

2

4

6

WTCD73 KO *

*

#

Control TAC-4W

bet

a-M

HC

0

2

4

6

**

Control TAC-4W

Myo

card

ial T

NF

a

0

2

4

6

**

#

*

Control TAC-4W

Myo

card

ial 3

-NT

0

2

4

6

*

*

#

*

B C D E

Figure S7. CD73 KO significantly exacerbates the TAC-induced increase of ventricular ANP, TNF and nitrotyrosine. TAC caused significant increases of ventricular myosin heavy chain (MHC) in both CD73 KO and wild type mice. CD73 KO tended to increase ventricular MHC after TAC, but the difference was not significant. *P<0.05 compared to the corresponding control; #p<0.05 compared to Wt-TAC.

LV

we

igh

t (m

g)

40

80

120

160

200ra

tio

of

LV

we

igh

t to

bo

dy

wei

gh

t(m

g/g

)

2

4

6

rati

o o

f L

V w

eig

ht

to

tib

ial l

eng

th(m

g/m

m)

2

4

6

8

10

Bo

dy

we

igh

t (g

)

10

20

30

40

* * *

Tib

ial l

en

gth

(m

m)

5

10

15

20

25 TAC TAC+CADO

n=

12

n=

19

n=

12

n=

19

n=

12

n=

19

n=

12

n=

19

n=

12

n=

19

Figure S8. The stable adenosine analogue CADO attenuated the TAC-induced ventricular hypertrophy as demonstrated by smaller increases of LV mass and ratio of LV mass to body weight. *P<0.05 compared to the vehicle treated group by Student t-test.

AControl

P-ERK

P-JNK

GAPDH

None CADOCADO

+ MRS1191 none CADOCADO

+ MRS1191

PE

EF

none PE

PE+CADO

PE+CADO+MRS 1191

p-E

RK

/GA

PD

H

0.2

0.4

0.6

0.8

1.0

1.2

1.4

none PE

PE+CADO

PE+CADO+MRS 1191

p-J

NK

/GA

PD

H

0.5

1.0

1.5

** B C

* * ***

Figure S9. Effect of CADO and A3R antagonist MRS1191 on PE-induced increases of p-JNK Thr183/Tyr185 and p-ERKThr202/Tyr204 expression. *P<0.05 between the indicated groups.