Download 395_2015_515_MOESM1_ESM - Springer Static Content Server

Regulatory Role of CARD3 in Left Ventricular Remodelling and Dysfunction
After Myocardial Infarction
Liangpeng Li • Xiaodi Wang • Wen Chen • Haoyu Qi • Ding-Sheng Jiang • Ling
Huang • Fuhua Huang • Liming Wang • Hongliang Li • Xin Chen
L. Li • X. Wang • W. Chen • H. Qi • F. Huang • L. Wang • X. Chen ()
Department of Thoracic and Cardiovascular Surgery, Nanjing Hospital Affiliated to
Nanjing Medical University, Changle Road 68, Nanjing 210006, Jiangsu, People’s
Republic of China.
D. Jiang • L. Huang • H. Li
Department of Cardiology, Renmin Hospital of Wuhan University, Wuhan 430060,
China; Cardiovascular Research Institute of Wuhan University, Wuhan 430060,
People’s Republic of China
Correspondence to
Xin Chen, PhD, MD
Professor and Director
Department of Thoracic and Cardiovascular Surgery
Nanjing Hospital Affiliated to Nanjing Medical University
Address: Changle Road 68, Nanjing 210006, Jiangsu, P. R. of China.
Tel & Fax: 86-25-52247821.
Email: [email protected]
Supplemental Tables
Supplementary Table 1. The primers for Real-Time PCR.
Primer name
Forward Primer
Reverse Primer
CARD3-human
TCCGCTGCTCGACAGTGAAAGA
GGGCAATTTCATGCAGGATGCGA
CARD3-mouse
AGGTTCAGAGCGCCCACCAATT
AAGGGCGTCCAGCGATTGGTTA
ANP-Mouse
ACCTGCTAGACCACCTGGAG
CCTTGGCTGTTATCTTCGGTACCGG
BNP-Mouse
GAGGTCACTCCTATCCTCTGG
GCCATTTCCTCCGACTTTTCTC
β-MHC-Mouse
CCGAGTCCCAGGTCAACAA
CTTCACGGGCACCCTTGGA
CTGF-Mouse
TGACCCCTGCGACCCACA
TACACCGACCCACCGAAGACACAG
Collagen I-Mouse
AGGCTTCAGTGGTTTGGATG
CACCAACAGCACCATCGTTA
Collagen III-Mouse
CCCAACCCAGAGATCCCATT
GAAGCACAGGAGCAGGTGTAGA
Bcl-2-Mouse
TGGTGGACAACATCGCCCTGTG
GGTCGCATGCTGGGGCCATATA
IL-1β-Mouse
CCGTGGACCTTCCAGGATGA
GGGAACGTCACACACCAGCA
MCP-1-Mouse
TGGCTCAGCCAGATGCAGT
CCAGCCTACTCATTGGGATCA
Supplemental Figures
Fig. S1 The sites of ligature were confirmed in KO and TG mouse hearts by Evans
blue staining. The heart sections of KO and TG mice after MI surgery are located in
the concordant ligation sites (n=5 mice per experimental group). A blue color
indicated normal myocardial tissue, while a pale gray color indicated infarcted tissue.
Fig. S2 The schematic diagram for the assessment of infarct size. The LV myocardial
midline was marked with the broken white line at the center between the epicardial
(red) and endocardial (green) surfaces and the length of the midline was measured as
midline circumference. Midline infarct length (blue arrow line) was taken as the
midline of the length of infarct.
Fig. S3 Immunohistochemical staining of the cardiac CARD3 protein in the heart
sections of wild type mice after sham or MI surgery. I: infarct area; B: border zone; R:
remote area.
Fig. S4 Effects of CARD3 on infarct size and cardiomyocyte apoptosis 3 days after MI.
a Histological analysis of hematoxylin and eosin (H&E)-stained WT and CARD3-KO
hearts at 3 days after MI (n=4 mice per experimental group). b H&E staining in NTG
and CARD3-TG mice at 3 days post-MI (n=4 mice per experimental group). c
Representative merged images via TUNEL staing showing cardiomyocyte apoptosis in
the border zone of WT and CARD3-KO hearts at 4 weeks post-MI (n=4 mice per
experimental group; magnification, 400 x). d TUNEL staining in the border zone of
NTG and CARD3-TG hearts at 4 weeks post-MI (n=4 mice per experimental group).