C57BL/6J (JAX® mice, stock #000664), mice were housed in an Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC) accredited facility at the University of Michigan Medical School. Mice were housed in a 12:12 light/dark cycle environment with controlled temperature (21–23°C) and humidity (30–70%). Mice were provided water ad libitum and were fed a phytoestrogen-reduced diet (16% protein, 4.0% fat, 48.5% carbohydrate, Teklad 2916 irradiated global rodent diet, Envigo) or a phytoestrogen-reduced, higher protein and fat diet (19% protein, 9% fat, 44.9% carbohydrate, Teklad 2919 irradiated global rodent diet, Envigo) for breeding and lactating females. Phytoestrogen-reduced diets were used to avoid any effects of exogenous dietary estrogens on AR expression in experimental mice. Adult male mice were single housed at least one week prior to euthanasia to control for housing status, which may impact testosterone levels 39 (link), and androgen-regulated AR expression 36 (link),37 (link). Adult female mice (group housed) were euthanized during diestrus, after completing at least two estrous cycles. Cycle stage was determined by vaginal lavage with predominately leukocytes 40 and confirmed by uterine weight below 100 mg 41 .
Sample size was 5–9 animals per sex and per age group (PND 12, PND 21, and adult).
Sample size was 5–9 animals per sex and per age group (PND 12, PND 21, and adult).