Nih 31

Animal studies were conducted under a protocol approved by the Animal Care and Use Committee of the National Cancer Institute. The generation of bi-transgenic mice harboring the CCSP-rtTA and TetO-regulated EGFR^L858R+T790M transgenes has been previously described [16 (link)]. To induce expression of mutant EGFR, mice were fed doxycycline-impregnated food pellets (NIH-31 with 650 ppm doxycycline, Harlan-Teklad, Madison, WI). Beginning at 6 weeks of age, groups of 3 mice received doxycycline (DOX) food or regular cereal diet. Mice were sacrified after 3 or 9 weeks. At sacrifice, lungs were removed and inflated with 10% neutral-buffered formalin. Lungs were embedded in paraffin and sectioned for immunohistochemistry.

Mouse studies and procedures were approved by the Eunice Kennedy Shriver National Institute of Child Health and Human Development Institute of Animal Care and Use Committee (IACUC). Fourteen adult male C57/Bl6 wild-type mice (20–26 wk old) were randomized to either the treatment group that was administered 10 mg/kg BW atorvastatin calcium (USP) in 0.9 % sterile saline or control group (0.9 % sterile saline) (n = 7/group). Treatment or vehicle was delivered by oral gavage (500 μl) for three consecutive days (09:00 h). Baseline, post-treatment, and follow-up blood samples were collected 1 week prior to, 24 h after, and 1 week after treatment (08:30–09:30 h). Group-housed mice (2–4/cage, 12-hour light/dark cycle, 23 °C) were provided ad libitum access to tap water and standard chow (NIH #31, Teklad) and acclimated to handling and restraint.

Adult male transgenic mice expressing a dominant-negative mutation in human PPARγ under the control of an endothelial-specific vascular cadherin promoter (E-V290M) was used as experimental models as reported.^{17 (link)} Age-and sex-matched non-transgenic (NT) littermates were used as controls. A separate study is currently ongoing specifically studying endothelial function in female E-V290M mice and in male and female offspring from E-V290M and NT mice undergoing arginine vasopressin-induced preeclampsia. NT and E-V290M mice were either fed TD.08290 (0.01–0.02% Na, Teklad) as LSD or standard chow (0.3% Na, NIH-31, Teklad) as normal salt diet. There was no difference in body or organ weights in response to diet or genotype (Table S1). Care of the mice met the standards set forth by the National Institutes of Health (NIH) guidelines for the care and use of experimental animals. All procedures were approved by The University of Iowa Animal Care and Use Committee.

Male and female (n = 10/group/time, 6 weeks of age at onset) CD1 mice (ICR:Crl) were housed in groups of 2–4, under a 12 h light cycle (lights on at 07:00 h) and provided with standard chow (REG; NIH-31, Harlan Teklad) and sterile water. All mice were fed standard chow for two weeks before introduction to the experimental diets. Mice were assigned to either ad libitum regular chow (REG, n = 40, 20 males) or a Standard American Diet (7 (link), 8 (link)) (SAD, n = 58, 29 males). At week 15, a subset of mice were switched from the SAD and onto the AID (n = 20, 10 males). All animals used in the present study were obtained, housed, cared for and used in accordance with the University of Alabama at Birmingham Institutional Animal Care and Use Committee guidelines.

WT C57BL/6 mice and FoxN1-Cre-transgenic mice were obtained from the Jackson Laboratory. Aire-deficient and Fezf2^flox/flox mice were gifts of Y. Takahama and N. Sestan, respectively (57 (link), 74 (link)). Fezf2^FoxN1-Cre mice were generated by crossing Fezf2^fl/fl and FoxN1-Cre mice. Mice were used between embryonic day 18.5 and 7 weeks of age, depending on the experiment. Embryonic mice were collected from timed pregnancies of 2- to 3-month-old females. Males were added to the cage and removed after detection of a vaginal plug (embryonic day 0.5). We did not detect any sex differences in 3- to 7-week mice and therefore pooled female and male mice for all analyses. Mice were housed on a 12-hour light/12-hour dark cycle, with ad libitum access to standard chow (NIH-31, Teklad). For experiments examining tissue steroid production, mice were collected between 10 a.m. and 3 p.m. (Zeitgeiber time 4 to 9). Sample sizes were selected on the basis of previous studies performed by the laboratory. Mouse genotypes were not randomized, and experimenters were not blinded. All protocols and procedures were approved by the National Cancer Institute (NCI) Animal Care and Use Committee (LICB-008) and followed the National Institutes of Health (NIH) Guide for the Care and Use of Laboratory Animals guidelines.