Myh11 cre

All the experiments were carried out according to the institutional guidelines and were approved by the Institutional Animal Care and Use Committee of University of California at Berkeley. MYH11-cre (#007742), Tie2-cre (#008863), Rosa-loxP-RFP (#007909) and Rosa-loxP-LacZ (#003474) mice were purchased from The Jackson Laboratory. Sox10-cre mice were a gift from Dr. Andrew S. McCallion and generated as described previously (Stine et al. Genesis, 2009, 47, 765–770). All the male MYH11-cre, Tie2-cre, and Sox10-cre mice were crossed with female Rosa-loxP-RFP or Rosa-loxP-lacZ mice to generate MYH11-cre/Rosa-loxP-RFP, Tie2-cre/Rosa-loxP-RFP, and Sox10-cre/Rosa-loxP-lacZ mice. PCR genotyping was performed according to the protocols provided by The Jackson Lab. The male transgenic mice of two months were used for experiments.

The BRD4 flox line was generated by Dr. Hill’s lab at University of Texas Southwestern [4 (link)], and was bred with male mice harboring a transgene of SMMHC-CreERT2 in the Y chromosome (Myh11Cre/+, The Jackson Laboratory: 019079, C57BL/6J strain) to generate male Myh11Cre/+: BRD4^flox/flox mice. Given that the Cre gene is inserted in the Y chromosome, only male mice express the Cre protein, and Cre-adenovirus induces KO of the BRD4 gene only in male mice. Mouse aortic rings with denuded endothelium from male Myh11Cre/+: BRD4^flox/flox mice were cultured with Ad-Null-GFP or Ad-Cre-GFP for 3 days to specifically knock out BRD4 in SMCs. Then, these tissues were used for myography. Before myography, vascular perfusion with a solution of sodium deoxycholate was used to remove the vascular endothelium.