CVR-2010-1022R2
Title page
Running title: Role of VPO1 in vascular pathogenesis of vascular diseases.
Ruizheng Shi
1*
, Changping Hu
2*
, Qiong Yuan
2
, Yongping Bai
1
, Tianlun Yang
1
, Jun
Peng
2
, Yuanjian Li
2
, Zehong Cao
3
,Guangjie Cheng
3#
, Guogang Zhang
1#
1
Department of Cardiovascular Medicine, Xiangya Hospital, Central South
University, Changsha 410008, China
2
Department of Pharmacology, School of Pharmaceutical Sciences,
Central South University, Changsha 410078, China
3 Division of Pulmonary, Allergy & Critical Care Medicine, Department of Medicine,
University of Alabama at Birmingham, Birmingham, AL 35294, USA
*
These two authors contributed equally to this work.
Correspondence to:
# Dr. Guogang Zhang, Department of Cardiovascular Medicine, Xiangya Hospital,
Central South University, Changsha 410008, China. E-mail: xyzgg2006@sina.com;
Phone: 086-731-84327695; Fax: 086-731-84327695.
Or Dr. Guangjie Cheng, Division of Pulmonary, Allergy & Critical Care Medicine,
Department of Medicine, University of Alabama at Birmingham, Birmingham, AL
35294, USA. E-mail: gjcheng@uab.edu; Phone: 1-205-975-8919; Fax:
1-205-935-8565.
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CVR-2010-1022R2
SUPPLEMENTARY FIGURE LEGENDS
Figure S1. Systolic blood pressure, plasma angiotensin II level and arterial
histological analysis in rats. A, Systolic blood pressure. B, Plasma angiotensin II level. C, Hematoxylin-eosin staining of thoracic aorta and mesenteric artery. D,
Statistical graph analysis of media thickness, lumen diameter, ratio of media thickness to lumen diameter, and mean nuclear area in artery media.
SHR: spontaneously hypertensive rats, n=10; WKY: Wistar Kyoto rats, n=10.
**
P < 0.01 vs SHR, magnification bar = 50 µm.
Figure S2. A larger portion of the gel for the VPO1 expression in aorta in SHRs.
Figure S3. Expression of MPO in arterial tissues in rats.
The immunohistochemistry analysis of MPO expression in thoracic aorta and mesenteric artery from SHR and
WKY. SHR: spontaneously hypertensive rats. WKY: Wistar Kyoto rats. n=10,
**
P <
0.01 vs SHR; magnification bar = 50 µm.
Figure S4. The efficiency of knockdown of VPO1 by shRNA. We used four shRNA
TargetSeqs against VPO1 to establish the VPO1-shRNA VSMCs and found that the second sequence had the best efficiency to inhibit the VPO1 expression. We therefore used the second shRNA TargetSeq to establish the VPO-shRNA VSMCs in the sequent experiments.
Figure S5. HOCl-mediated A10 VSMCs proliferation. A, BrdU incorporation. B,
Cell cycle analysis by flow cytometry . Control: wild-type cells; HOCl: wild-type cells
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CVR-2010-1022R2 treated with 20 µmol/L HOCl for 1 h, followed by washing and continued incubation in DMEM for up to 24 h ; VPO1-shRNA: cells transfected with VPO1-shRNA;
VPO1-shRNA+HOCl: VPO1-shRNA tranfected cells treated with 20 µmol/L HOCl for 1 h, followed by washing and continued incubation in DMEM for up to 24 h .
**
P <
0.01 vs Control,
++
P < 0.01 vs HOCl. n=3. Data are representatives of three independent experiments.
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CVR-2010-1022R2
Figure S1
A
220
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B
**
WKY
SHR
** ** ** ** ** ** ** **
12 14 16 18 20
Time (Week)
600
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0
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SHR
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CVR-2010-1022R2
C
D
D
2.5
2.0
1.5
1.0
.5
**
WKY
SHR
0.0
Mesenteric artery Thoracic aorta
.16
.14
.12
.10
WKY
SHR
**
.08
.06
**
.04
.02
0.00
Mesenteric artery Thoracic aorta
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SHR
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Mesenteric artery Thoracic aorta
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SHR
**
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10
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Mesenteric artery Thoracic aorta
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CVR-2010-1022R2
Figure S2
Figure S5
50
40
35
50
140
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70
37
25
20
KD
250
WKY SHR WKY SHR
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CVR-2010-1022R2
Figure S3
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CVR-2010-1022R2
VPO1
-actin
1 2
3 4
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CVR-2010-1022R2
Figure S5
A 200
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** ++
B
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9