RNA-seq analysis of lung smooth muscle cells

Lung smooth muscle cells are including bronchiolar and vascular smooth muscle cells. In order to get adult lung smooth muscle cells, we use transgenic mouse line with smooth muscle actin creERT2, which is a transgenic cre recombinase in Acta2 (contractile smooth muscle cell gene). This mouse line also contains a CAG promoter-driven red fluorescent protein variant (tdTomato) - all inserted into the ROSA26 locus. This mouse line express robust tdTomato fluorescence following cre-mediated recombination after Tamoxifen injection. Tamoxifen (Sigma) was administered intraperitoneally with 75mg/kg body weight daily for 10 days to 8 weeks old transgenic mice. After 2 months, mice were sacrificed and lung tissue was dissected from mice and dissociated into single cells by enzyme digestion. Then, cell sorting was done with FACSAria (BD Biosciences).