Tlr4 ko mice

TLR4 KO mice were purchased from Jackson Laboratory (Bar Harbor, ME, USA). The control mice (C57BL/6J) were provided by the Laboratory Animal Center of Central South University. All mice used in this study were male, 8–10 weeks of age and weighed between 25 g and 30 g. Animals were maintained in a 12-h light/dark cycle. The Animal Care and Use Committee of the Central South University approved all animal protocols (NO.201403122). All experiments were conducted in accordance with the National Institutes of Health Guidelines for the Care and Use of Laboratory Animals.

5-6 weeks of age, female C57BL/6 TLR2 knockout (KO) mice, TLR4 KO mice, C57BL/6 OT-I mice, and OT-II T TCR transgenic mice were purchased from the Jackson Laboratory (Bar Harbor, ME, USA). All animals were maintained under specific pathogen-free (SPF) barrier conditions at Preclinical Research Center (PCRC) of Chungnam National University Hospital, Daejeon, Korea. All mice used are in accordance with the guide for care and used of the Korean Food and Drug Administration (KFDA). All animal studies used were licensed by the Ethics Committee and Institutional Animal Care and Use Committee (Permit number 2014-0197-3) of the Laboratory Animal Research Center at IACUC (CNU-00284) of Chungnam National University (Daejeon, Korea).

CTRP1-KO mice were purchased from Cyagen Biosciences (Guangzhou, China, KOCMP-07979). C57BL6J mice (male) were purchased from Beijing Huafukang Biology Co., Ltd (Beijing, China). TLR4-KO mice were purchased from Jackson Laboratory (stock no: 029015). C57BL6J mice purchased from the Chinese Academy of Medical Sciences (Beijing, China) were subjected to intraperitoneal injection of recombinant human CTRP1 full-length protein (Abcam, ab151376; 200 μg/kg) every other day from days 7 to 28 post MI). For the macrophage clearance experiment, mice were subjected to intravenous injection of clodronate liposomes (CLs; 150 mL; 5 mg/mL; Liposoma, The Netherlands) at 10- and 14-days post MI. For the TLR4 deficiency experiment, TLR4-KO mice were subjected to intraperitoneal injection of recombinant human CTRP1 (200 μg/kg, from 7 days to 28 days post MI). All animal experiments were approved by the Institutional Animal Care and Use Committee of Huai’an First People’s Hospital, Nanjing Medical University (Huai’an, China).