Alzet osmotic mini pumps model 2002

Male C57B6 mice, aged 8–10 weeks, were subjected to the AB operation for two weeks to produce pressure overload [27 (link)]. Briefly, after anesthetization (ketamine, 25 mg/kg/IP, Sigma, Shanghai, China) and artificial ventilation, the transverse aorta was banded with a 7-0 nylon suture by binding the aorta together with a blunted 26-gauge needle, which was pulled out later. AG-490 (1 mg/kg body weight per day), TSA (0.6 mg/kg/day) or PBS (vehicle) was continuously administered by Alzet osmotic mini-pumps (Model 2002, DURECT, Cupertino, CA, USA) implanted subcutaneously into the back of mice from 3 days before AB until AB was finished. Animals recovered for 2 weeks after AB to develop cardiac hypertrophy. All animals were maintained in accordance with the guidelines of the National Institute of Health (NIH) (Guide for the Care and Use of Laboratory Animals, 1996), the European Communities Council. The protocol is approved by Animal Care and Use Committee of all authors’ institutions (approved at 10 November 2011, identification code: SHXQYY-100015, contact name of institutional review board: Shen Wang).

Pressure overload was produced by the transverse aorta constriction (TAC) as previously described [20 (link)]. C57BL/6 mice aged 8–10 weeks were subjected to TAC or sham operation after anaesthetized with ketamine (25 mg/kg IP). Briefly, after anaesthesia and artificial ventilation, the transverse aorta was constricted with the 7-0 nylon suture by ligating the aorta together with a blunted 27-gauge needle, which was pulled out later. From 2 to 4 weeks after TAC, urantide (30 μg/kg/day, Peptides International Inc., Louisville, KY, USA) or vehicle were respectively continuously administered by Alzet osmotic minipumps (Model 2002; DURECT, Cupertino, CA, USA) implanted subcutaneously into the back of mice. SP600125 or Dimethyl sulfoxide (DMSO) were injected intraperitoneally either with 40 mg/kg bodyweight every 3 days for 2 weeks. At 4 week after TAC, the number of CSPs was determined. All animal experimental protocols were approved by the Animal Care and Use Committee of Fudan University and in compliance with ‘Guidelines for the Care and Use of Laboratory Animals’ published by the National Academy Press (NIH Publication No. 85–23, revised 1996).