Sterile C-section and fostering to conventional Swiss Webster recipient females

Evening-morning timed matings to generate donor susceptible or resistant females yielding iECre;Krit1^fl/fl or iECre;Ccm2^fl/fl pups were performed and timed pregnant Swiss Webster females (Charles River 024) were ordered to serve as foster mothers. To prevent delivery of the pups, at E16.5, donor females were injected subcutaneously with 100 μL of a 15 μg/mL solution of medroxyprogesterone (Sigma Aldrich, M1629) dissolved in DMSO. The morning of E19.5, the donor mother was euthanized by cervical dislocation and submerged in a warm sterile solution of 1% VirkonS^TM/PBS (weight/volume) for one minute. The uterus was then dissected in a sterile laminar flow hood, submerged in a warm sterile solution of 1% VirkonS^TM/PBS for one minute and quickly rinsed with warm sterile PBS. Pups were then removed from the uterus and fostered to the Swiss Webster recipient female. The following morning, induction of the neonatal CCM model was performed as described above.