C57bl 6ntac females

In vitro fertilization was performed using FERTIUP Mouse Preincubation Medium and CARD MEDIA (Kyudo Company) according to the manufacturer’s protocol. Non-virgin Chd2^RH/RH, Mecp2^del/Y, and A/J (The Jackson Laboratory, Stock No. 000646) males that were >8 weeks old were used as sperm donors. For experiments using Chd2 or Mecp2 sperm, oocytes from C57Bl/6NTac females (Taconic) were used. For experiments using A/J sperm, oocytes from C57Bl/6J females (The Jackson Laboratory, Stock No. 000664) were used. Following IVF, embryos were cultured for ~8 hours and then injected using the PNI protocol described below.

C57BL/6NTac females were purchased from Taconic and housed on a 12h light:12h dark-cycle with free access to chow diet (Altromin # 1324, Chr. Pedersen, Denmark) and tap water. The mice arrived at age 10-12 weeks and were acclimatized for at least 1 week before experimentation. Wild type and muscle-specific dominant-negative kinase-dead α2 (KD) AMPK female mice were provided by Morris Birnbaum [30 (link)] and have been backcrossed at our facility for >10 generations onto the C57BL/6NTac strain. All experiments were carried out on 11-14 wk. old mice. All experiments were approved by the Danish Animal Experiments Inspectorate and carried out in accordance with the “European Convention for the Protection of Vertebrate Animals Used for Experiments and Other Scientific Purposes”.