Rosa26 cag loxp stop loxp tdtomato

Atoh1-CreER™ (Chow et al., 2006 (link)) and Prestin^CreERT2(Fang et al., 2012 (link)) mice were obtained from Dr. Suzanne Baker and Dr. Jian Zuo at St. Jude Children's Research Hospital (Memphis, TN), respectively. Pou4f3^loxP/loxP (stock # 10560) (Badea and Nathans, 2011 (link)) and ROSA26^{CAG−loxP−stop−loxP−tdTomato} (ROSA26^tdTomato) mice (stock # 7914; also called Ai14) (Madisen et al., 2010 (link)) were purchased from The Jackson Laboratory (Bar Harbor, ME). All genotyping was performed by Transnetyx, Inc. (Cordova, TN) and mice of both genders were used in the study. All animal work was performed in accordance with approved animal protocols from the Institutional Animal Care and Use Committee at Southern Illinois University School of Medicine.

TRE-H2B-EGFP (Foudi et al., 2009 (link)) knock-in, Col2a1-creER transgenic (Nakamura et al., 2006 (link)), Pthlh-LacZ/null knock-in (Chen et al., 2006 (link)), Gas1-LacZ/null knock-in (Martinelli and Fan, 2007 (link)), Pthlh-creER transgenic (Mizuhashi et al., 2018 (link)) mice have been described elsewhere. Rosa26-CAG-loxP-stop-loxP-tdTomato (Ai14: R26R-tdTomato, JAX007914), Apc-floxed (JAX009045) mice (Cheung et al., 2010 (link)) were acquired from the Jackson Laboratory. All procedures were conducted in compliance with the Guidelines for the Care and Use of Laboratory Animals approved by the University Michigan’s Institutional Animal Care and Use Committee (IACUC), protocol 7681 and 9496. All mice were housed in a specific pathogen-free condition, and analyzed in a mixed background. Mice were identified by micro-tattooing or ear tags. Tail biopsies of mice were lysed by a HotShot protocol (incubating the tail sample at 95°C for 30 min in an alkaline lysis reagent followed by neutralization) and used for GoTaq Green Master Mix PCR-based genotyping (Promega, and Nexus X2, Madison, WI). Mice were euthanized by over-dosage of carbon dioxide or decapitation under inhalation anesthesia in a drop jar (Fluriso, Isoflurane USP, VetOne, Boise, ID).

Shhgfpcre (JAX stock #05622), Piezo1flox (JAX stock #029213), Piezo2flox (JAX stock #027720) and Rosa26-CAG-loxp-stop-loxp-tdTomato (JAX stock #007914) were obtained from Jackson Laboratory. Data reported herein have been compiled from the examination of multiple embryonic and postnatal time points for wildtype (WT), heterozygous and conditional knockout animals. Both male and female mice were used in this study; experimental comparisons that underwent naphthalene injury controlled for sex. Pregnant dams and adults were killed for experiments through CO2 asphyxiation and cervical dislocation followed by isolation of vital postnatal organs and embryos. Fetal and neonate mice (< 10 days) were put on ice until motionless and euthanized via decapitation. Timed pregnancies were confirmed through visualization of vaginal plugs with noon on the day plugs were detected designated as E0.5. Mutant and control animals were studied at E15.5, E18.5, P0, P15 and 6 weeks of age. Mice were bred and housed in the University of Wisconsin-Madison Biomedical Research Model Services Laboratory. Animal procedures were approved by the University of Wisconsin-Madison Institutional Animal Care and Use Committee and conducted in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals [55 ].