Animals and experimental design: Treatment of SOD1G93A mice
We used the hybrid strain of SOD1G93A mouse as an ALS model (B6SJL-Tg[SOD1-G93A]1Gur). Non-transgenic wild-type (WT) littermates were used as controls. The animals used were from colonies that had been bred in different animal facilities for years, as specified below. The transgenic offspring was identified by polymerase chain reaction (PCR) amplification of DNA extracted from the tail, to identify hemizygous transgenic mice and WT. The colony was maintained under stander conditions (food and water ad libitum, room temperature of 22 ± 2 °C and 12:12-h light–dark cycle) at the Animal Service of the UAB and at the animal facilities in Centro de Investigación Biomédica de Aragón University of Zaragoza. Mice were handled in accordance with the guidelines of the European Union Council (Directive 2010/63/EU) and Spanish regulations (RD53/2013) on the use of laboratory animals. All experimental procedures were approved by the Ethics Committee of the Universitat Autònoma de Barcelona (CEEAH-UAB: 2969 and 4273) and by the Ethic Committee for Animal Experiments from the University of Zaragoza (PI45/18, PI15/21, PI51/21, PI02/23). Humane endpoint criterium in the survival studies was considered when animals lost the righting reflex for longer than 30 s.