The Pax9-CreER founder mouse (F0) was mated with Bl6 females to generate F1 Pax9-CreER mice. ROSA26LoxP−STOP−LoxP−tdTomato (tdTomato reporter) mice were obtained from the Jackson Laboratory (JAX no. 007905) (Madisen et al., 2010 (link)). tdTomato reporter female mice were mated with Pax9-CreER F1 male mice and received 1.5mg/10g bodyweight tamoxifen (Sigma T5648) at E8.5 through E14.5. The female mice were euthanized 48 hours after injection to collect Pax9-CreER;tdTomato embryos. Adult Pax9-CreER;tdTomato mice received 5 daily injections of 1.5 mg/10 g bodyweight tamoxifen and were euthanized 48 hours after the last injection. Whole samples were imaged using a Leica MZ10F. For cryosections, samples at various stages were fixed in 4% paraformaldehyde, passed through a sucrose series, then embedded in optimal cutting temperature (OCT) compound (Tissue-Tek, Sakura) and frozen onto a dry ice block to solidify. Embedded samples were cryosectioned at 7μm thickness using a cryostat (Leica CM1850). Sections were counterstained with DAPI (Sigma, D9542). Images were captured using a fluorescence microscope (Leica DMI 3000B) with filter settings for DAPI/FITC/TRITC.
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Generating Pax9-CreER;tdTomato Reporter Mice
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Corresponding Organization : USC Norris Comprehensive Cancer Center
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Variable analysis
independent variables
- Tamoxifen administration at E8.5 through E14.5
- Tamoxifen administration for 5 daily injections in adult mice
- Collection of Pax9-CreER;tdTomato embryos 48 hours after injection
- Euthanasia of adult Pax9-CreER;tdTomato mice 48 hours after the last injection
- Mating of Pax9-CreER founder mouse (F0) with Bl6 females to generate F1 Pax9-CreER mice
- Use of ROSA26LoxP−STOP−LoxP−tdTomato (tdTomato reporter) mice
- Mating of tdTomato reporter female mice with Pax9-CreER F1 male mice
- Fixation of samples in 4% paraformaldehyde, passage through a sucrose series, embedding in OCT compound, and cryosectioning at 7μm thickness
- Counterstaining of sections with DAPI
- Imaging using a fluorescence microscope with filter settings for DAPI/FITC/TRITC
- None specified
- None specified
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