Myh11 creert2

Animal experiments were performed according to procedures approved by the Dana-Farber Cancer Institute IACUC. Myh11-Cre (stock #007742), Myh11-Cre/ERT2 (stock #019079), tdTomato reporter mice (stock #007914), and ROSA^mT/mG reporter mice (stock #007676) were obtained from Jackson Laboratories. The generation of TRAP and Ucp1-Cre transgenic mice is described in detail in the Supplemental Experimental Procedures. UCP1-TRAP breeding pairs were housed at 30°C and litters were weaned to room temperature at 3 weeks. Unless otherwise stated, for cold exposure experiments 4 week-old female UCP1-TRAP mice were individually housed at 4°C for two weeks.

Lmo7^−/− mice in C57BL/6J background were obtained from Dr. Jun Miyoshi (Osaka Medical Center, Japan)^{11 (link)}. Experimental Lmo7^−/− and littermate control mice were generated from heterozygous knockout breeding pairs. Lmo7^fl/fl mice obtained from Dr. Ju Chen (UCSD)^{14 (link)} were originally in Black Swiss background and backcrossed to BL6 for over six generations. Female Lmo7^fl/fl mice were crossed with male Myh11-CreER^T2 (on Y-chromosome, Jackson Laboratory) mice to generate female Lmo7^fl/+ and male Lmo7^fl/+/Myh11-CreER^T2, which were intercrossed to generate male Lmo7^fl/fl/Myh11-CreER^T2 or Lmo7^+/+/Myh11-CreER^T2. Male Lmo7^fl/fl/Myh11-CreER^T2 mice were then crossed with littermate female Lmo7^fl/fl to generate experimental Lmo7^fl/fl/Myh11-CreER^T2 mice, and Lmo7^+/+/Myh11-CreER^T2 were crossed with littermate female Lmo7^+/+ to generate Lmo7^+/+/Myh11-CreER^T2 (control) mice. Experimental and control mice were age- and size-matched. Lmo7 deletion was induced by injecting 6-week-old mice with 50 mg/kg tamoxifen for 5 days, followed by 5 day recovery prior to surgery. Control mice also received tamoxifen. Genotyping was performed by PCR. Mice were housed in pathogen-free conditions in Yale University animal facilities. All experiments were approved by the Yale University Institutional Animal Care and Use Committee.

SMC-specific Cre mice (Myh11-Cre-ER^T2) were purchased from Jackson Laboratory. As Myh11-Cre-ER^T2 was inserted into Y chromosome, only male mice were used.^{45 (link)} By crossing male Myh11-Cre-ER^T2 mice with female FAK flox/flox (FAK^FL/FL) mice, we generated FAK^FL/FL;Myh11-Cre-ER^T2 mice. Male FAK^FL/FL;Myh11-Cre-ER^T2 mice were crossed with female FAK wild-type (WT)/kinase-dead (KD) (FAK^WT/KD) mice to generate FAK^FL/WT;Myh11-Cre-ER^T2 and FAK^FL/KD;Myh11-Cre-ER^T2 mice (Online Figure I). The flox FAK allele was excised by treating male mice (6-weeks-old) with tamoxifen (75 mg/kg, intraperitoneal, every other day) for 2 weeks, generating FAK^−/WT;Myh11-Cre-ER^T2 (FAK-WT) and FAK^−/KD;Myh11-Cre-ER^T2 (FAK-KD) mice.^{46 (link)}

Pdgfra^H2B-eGFP (Jackson Laboratories (JAX) strain 007669)^{42 (link)}, Rosa26R^LSL-TdTomato (JAX strain 007909), Myh11^Cre(ER-T2) (JAX strain 019079), Pdgfra^Cre(ER-T2) (JAX strain 032770), Etv1^Cre(ER-T2) (JAX strain 013048) and Rosa26R^LSL-DTA (JAX strain 009669) mouse lines were purchased from Jackson Laboratories (Table S1). Adult mice were more than 8 weeks of age at the time of treatments or cell isolations. Animals were housed in a specific pathogen-free barrier facility, maintained on a 12-hour light/dark cycle, and had ad libitum access to standard chow and water. All experiments used mice of both sexes and littermates as controls. All animal procedures and experiments were approved and monitored by the Animal Care and Use Committee at the Dana-Farber Cancer Institute.

All mice were housed and treated in accordance with the IACUC protocol approved at the University of California, San Francisco. Mice between the ages of 8-12 weeks old were used for the experiments and littermates were used as controls with balance of gender between groups. C57BL/6 mice were obtained from Jackson Laboratory. The generation and genotyping of the Gli1^creERT2, R26R^EYFP, R26R^tdTomato, R26R^DTR, Ubc^creERT2 and Myh11^creERT2 lines were performed as previously described by The Jackson Laboratory. Generation and genotyping of the Sftpc^{creERT246 (link)} and Yeti^{YFP27 (link)} lines were performed as previously described. The Hhip^flox/flox line was generated in our lab, and genotyping primers are listed in KEY RESOURCES TABLE.