All animal procedures were approved by the Institutional Animal Care and Use (IACUCs) Committees at the University of Pittsburgh and Magee-Womens Research Institute (protocol #22010530) in compliance with the National Institute of Health’s Office of Laboratory Animal Welfare Guide for the Care and Use of Laboratory Animals and the ARRIVE guidelines. CB6-Tg (CAG-EGFP/CETN2)3-4Jgg/J mice (Stock number: 00823445) were obtained from the Jackson Laboratory (Bar Harbor, ME) as juveniles, bred, and analyzed as described previously15 (link),17 (link). All mice were housed in an Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC)-accredited mouse facility and tissues collected after humanely euthanizing mice by carbon dioxide (CO2) asphyxiation (55–60 cubic feet per hour flow rate; 2–4 min) followed by cervical dislocation using approved methods by the American Veterinary Medical Association (AVMA) and our University-approved IACUC protocols. Tissues were harvested within 5 min post-euthanization. We investigated isolated spermatogenic cells from 54 testes from 27 male mice and produced 983 spermatogenic cells for analysis.
GFP expression determination by PCR. Genomic DNA was isolated for GFP detection in GFP CETN2-expressing mice, using tail tip tissues (< 5 mm) and PCR with MyTaq Extract-PCR Kit (Bioline, Taunton, MA) as previously described17 (link).
GFP expression determination by PCR. Genomic DNA was isolated for GFP detection in GFP CETN2-expressing mice, using tail tip tissues (< 5 mm) and PCR with MyTaq Extract-PCR Kit (Bioline, Taunton, MA) as previously described17 (link).
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