Cd1 icr mice

GST pull-down assays were performed as described previously [38 (link)]. In brief, adult male ICR (CD-1) mice were purchased from Charles River Laboratory and anaesthetized with the help of Isoflurane (Hospira Inc, IL). Brains were removed quickly, homogenized in the ice-cold lysis buffer containing 50 mM HEPES, pH 7.4, 100 mM NaCl, 2 mM EDTA, 1 % Triton X-100 supplemented with protease and phosphatase inhibitors cocktails (Roche Life Science, Indianapolis, IN). After removal of the insoluble fractions, soluble supernatant was incubated at 4 °C with equal amount of GST-tagged recombinant purified proteins coupled with glutathione resin. Samples were washed, eluted out and separated on one-dimensional gel electrophoresis using 4-12 % Bis-Tris Gel (Life technologies, Grand Island, NY). Gels were then subjected to Colloidal Blue staining and the excised bands were subjected to mass spectrometry-based analysis.

Male and female ICR CD-1 mice at 7–9 months old were employed for all experiments (Charles River, Sulzfeld, Germany). Mice were housed in groups of four in the animal facilities of the Max Planck Institute of Psychiatry in Munich, Germany, from weaning and were maintained under standard conditions (12L:12D light cycle, lights on at 07:00 AM, temperature 23 ± 2°C) with food and water available ad libitum. All experiments were approved by and conducted in accordance with the regulations of the local Animal Care and Use Committee (Government of Upper Bavaria, Munich, Germany), under licenses Az.: 55.2-1-54-2532-148-2012, Az.:55.2-1-54-2532-32-2016 and ROB-55.2–2532.Vet_02-18-50. The fur of all mice was marked using four different colors under mild isoflurane anesthesia and mice were left to recover for several days before the start of the experiment. On day 1, animals were transferred to the SB (see ‘The ‘Social Box’ paradigm’ section), for a total of 5 days (five light periods and five dark periods). On day 6, animals were removed from the SB and placed in their original cage under standard housing conditions for the rest of the experimental procedure (see ‘Chronic mild stress protocol’ and ‘Behavioral battery’ sections).

The mice with cardiomyocyte‐specific Gata4 deletion (CM‐G4‐KO: Tg(β‐MHC‐Cre);Gata4^flox/flox) were previously described and were maintained on a mixed SV129/CD1 background (Oka et al, 2006; Heineke et al, 2007). Littermate Gata4^flox/flox mice were used as control mice as described before (Oka et al, 2006; Heineke et al, 2007). Wild‐type mice (WT) with and without the β‐MHC‐Cre transgene (WT‐Cre; Tg(β‐MHC‐Cre) were kept on the same mixed SV129/CD1 background. ICR‐CD1 mice were obtained from Charles River Laboratories. Male and female neonatal mice were equally used throughout the study. The animals had free access to water and a standard diet and were maintained on a 12‐h light and dark cycle at a room temperature of 22 ± 2°C. The number and age of mice used is indicated in each figure. All animal procedures described in this study were approved by the local state authorities (the Lower Saxony State Office for Consumer Protection and Food Safety, Germany, file number: 33.12‐42502‐04‐11/0488).