PDGFRβ-CreERT2 transgenic mice expressing a tamoxifen-inducible Cre recombinase under the endogenous PDGFRβ promoter (B6.Cg-Tg(Pdgfrb-cre/ERT2)6096Rha/J, The Jackson Laboratory) were crossed with Tdtomato reporter mice Ai14 (B6.Cg-Gt(ROSA)26Sortm14(CAG-tdTomato)Hze/J, The Jackson Laboratory) to generate the bitransgenic reporter mice PDGFRβ-CreERT2;Ai14. Reporter mice were fed a tamoxifen containing diet postnatally between weeks 3–7. Cells expressing PDGFRβ during tamoxifen exposure and their daughter cells were identified by permanent expression of the red fluorescent protein Tdtomato. Tamoxifen-exposed PDGFRβ-CreERT2;Ai14 mice underwent lung injury by rhFasL/mechanical ventilation as described above. Harvested lungs were inflated with 10% normal buffered formalin (Sigma) at 25cm H2O column pressure, fixed in 10% normal buffered formalin overnight and stored in 70% EtOH until processing for paraffin embedding. Fluorescently labeled RNA probes against Angptl4 and Tdtomato were purchased from Advanced Cell Diagnostics, Inc. Hybridization of RNA targets in paraffin-embedded lung sections using the RNAScope® 2.0 Assay (Advanced Cell Diagnostics, Inc.) was performed per manufacturer’s protocol (16 ).
B6 cg tg pdgfrb cre ert2 6096rha j
Manufactured by Jackson ImmunoResearch
The B6.Cg-Tg(Pdgfrb-cre/ERT2)6096Rha/J mouse strain is a genetically modified mouse model that expresses a tamoxifen-inducible Cre recombinase under the control of the platelet-derived growth factor receptor beta (Pdgfrb) promoter. This strain can be used to achieve cell type-specific gene manipulation in cells expressing Pdgfrb.
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2 protocols using b6 cg tg pdgfrb cre ert2 6096rha j
1
Lineage Tracing of PDGFRβ-Expressing Cells in Lung Injury
2
Conditional Knockout of Gnaq and Gna11 in Mice
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For the germline mouse studies, mice with floxed alleles for Gnaq and germline deficiency in Gna11 (Gnaqfl/fl;Gna11−/−) were crossed with mice that express Cre recombinase under the control of the Pdgfrb gene (Pdgfrb-Cre+/−). Pdgfrb-Cre+/−;Gnaq+/fl;Gna11+/− offspring from this F1 generation were then bred with Gnaqfl/fl;Gna11−/− founders to produce an F2 generation, including Pdgfrb-Cre+/−;Gnaqfl/fl;Gna11−/− mice. The genetic background for all mice was predominantly C57BL6, with a minimum of a six backcross generations. The generation of Gnaqfl/fl;Gna11−/− and Pdgfrb-Cre+/− mice has been described previously (Foo et al., 2006 (link); Offermanns et al., 1998 (link); Wettschureck et al., 2001 (link)).
For the tamoxifen-inducible mouse gene knockout studies, the same breeding strategy was used as for the germline studies but substituting Pdgfrb-Cre/ERT2+/− mice [029684, B6.Cg-Tg(Pdgfrb-cre/ERT2)6096Rha/J, The Jackson Laboratory] for Pdgfrb-Cre+/− animals. The generation of these mice has been described elsewhere (Gerl et al., 2015 (link)).
For the tamoxifen-inducible mouse gene knockout studies, the same breeding strategy was used as for the germline studies but substituting Pdgfrb-Cre/ERT2+/− mice [029684, B6.Cg-Tg(Pdgfrb-cre/ERT2)6096Rha/J, The Jackson Laboratory] for Pdgfrb-Cre+/− animals. The generation of these mice has been described elsewhere (Gerl et al., 2015 (link)).
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