Regulatory Role ofCARD3in Left Ventricular Remodellingand DysfunctionAfter 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:

Supplemental Tables

SupplementaryTable 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

SupplementalFigures

Fig. S1 The sites of ligaturewere 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 Theschematic 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. S3Immunohistochemical staining of the cardiac CARD3 proteinin 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 CARD3on infarct size and cardiomyocyte apoptosis 3days after MI. a Histological analysis of hematoxylin and eosin (H&E)-stained WT and CARD3-KO hearts at 3days after MI(n=4 mice per experimental group). b H&E staining in NTG and CARD3-TGmice at 3days post-MI (n=4 mice per experimental group). c Representative merged images via TUNEL staing showing cardiomyocyte apoptosis in theborder zoneof 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).