C57bl 6j wild type

C57BL/6 J wild-type (Charles River Laboratories) and Igfbpl1^−/− mice at a C57BL/6 J genetic background (Knockout Mouse Project Repository, University of California at Davis) were used in these experiments. All mouse studies were approved by the Schepens Eye Research Institute Animal Care and Use Committee and performed in accordance with institutional and federal guidelines.

All experimental procedures involving the use of animals were carried out in accordance with EU Directive 2010/63/EU and under approval of the EMBL Animal Use Committee and Italian Ministry of Health License 541/2015-PR to C.G. Animals were maintained in a temperature and humidity controlled environment with food and water provided ad libitum and 12 hr/12 hr light-dark cycle with lights on at 7:00. Experimental male C57BL/6J wild type (Charles River) and Esr1::Cre (Stock No. 017913, Jackson Laboratory) mice were switched to reverse dark-light cycle (lights off at 9:00) and singly housed in the home cage of the behavioral apparatus at least 1 week before initiating the experimental procedures and tested during the dark period under red lighting (two 1W LED lights). The custom Plexiglas behavioral apparatus consisted of three detachable parts: 1) home cage with dimensions 25 × 25×25 cm with a Y-shaped slit, 4 cm wide at the bottom serving as an entrance closed by sliding doors, 2) stimulus chamber identical to home cage, and 3) a corridor 46 × 12×30 cm connecting home cage and stimulus chamber (modified from Silva et al., 2013 (link)). Aggressor mice were CD-1 adult retired breeders (Charles River) screened for robust aggression and singly housed (Franklin et al., 2017 (link)). Subordinate mice were 9–15 week old BALB/c males bred at EMBL. Mice cages were changed weekly.

All experiments were performed using 8–12 weeks old C57Bl/6J wild-type (WT, in total 20 animals obtained from Charles River Laboratories) mice of both sexes. All animals were housed under standard laboratory conditions according to a normal day/night rhythm with ad libitum access to food and water. For anaesthesia, a mixture of xylazine (10 mg/kg body weight) and ketamine (70 mg/kg body weight, Anesketin) was i.p. administered. Additionally, for surgery and intravitreal (ivt) injections, a drop of local analgesic, oxybuprocaine hydrochloride (0.4%, Unicaïne, Théa, Wetteren, Belgium) was applied to the treated eye. During recovery from anaesthesia, an ocular ointment (Tobrex; Alcon, Puurs, Belgium) was applied on the cornea to prevent corneal desiccation. All animals were sacrificed with an i.p. injection of an overdose of sodium pentobarbital (30 mg/kg, Nembutal).
All animal experiments were approved by the Institutional Ethical Committee of KU Leuven (P260/2014) and followed the guidelines of the Association for Research in Vision and Ophthalmology and the European Communities Council Directive of 22 September 2010 (2010/63/EU).

Female C57Bl/6J (wild-type) and C57Bl/6J-ob/ob mice were obtained from Charles River (Sulzfeld, Germany). At the age of 12 weeks, mice were caged individually using cages with an enriched environment. Prior to wounding, mice were randomly assigned to different experimental groups, monitored for body weight, and wounded as described below. The animal experiments were performed according to the guidelines and approval of the local Ethics Animal Review Board (Regierungspräsidium Darmstadt, D-64278 Darmstadt, Germany). The approval number to this project was V54-19c20/15 –F143/10).

Wild-type C57BL/6 J (Charles River Laboratories, UK) and heterozygous Ins2^Akita (The Jackson Laboratory, USA) male mice were used for experiments. Ins2^Akita, hereafter called Akita, carry a single nucleotide substitution in the insulin 2 gene (Ins2). This mutation results in misfolding of proinsulin 2 and eventually leads to ER stress and pancreatic β cell failure and ultimately to severe hyperglycaemia. At 5 weeks of age, Akita mice developed hyperglycaemia with non-fasting blood glucose levels higher than 25 mM. For chronic MitoGamide treatment, 6-week-old mice were subjected to daily oral administration via gavage of MitoGamide (10 mg/kg) or vehicle (sterile water) for 12 weeks. Blood glucose and HbA1c levels (service provided by Core Biochemical Assay Laboratory, Cambridge University Hospital) were monitored during the treatment period. Cardiac function and morphology were evaluated using echocardiography (Vevo 3100, VisualSonics). The parasternal long-axis view (B mode), the short-axis view (M mode) and blood flow velocity (PW mode) at mitral valve were obtained, and measurements of cardiac structure and function were accessed using Vevo Lab (VisualSonics) in a completely blinded manner.

Wild type C57Bl/6J and Balb/cAnNCrl male mice of 8–10 weeks old were obtained from Charles River (Maastricht, the Netherlands). Mice transgenic for human Interleukin-37 (hIL-37Tg) were created as described previously [5 (link)] and bred and housed under specific pathogen-free conditions in the animal facility of the Radboud university medical center in Nijmegen. Transgenic expression of IL-37 was confirmed by standard genotyping protocols on ear tissue samples. All mice received water and food ad libitum, and only age- and sex-matched mice were included in experiments. All animal experiments were carried out following the guidelines of local and national Animal Ethics Committees and after permission granted by the Animal Ethics Committee of the Radboud University Nijmegen (Permit Number 2011–024). Mice were sacrificed by cervical dislocation.