Comparative ATAC-seq of cultured mouse Acta2-R179C aortic SMCs versus healthy controls

Explanted aortic SMCs from transgenic,Acta2-R179C/+ mice and littermate controls were cultured and subjected to assay for transposase accessible chromatin by sequencing (ATAC-seq) to determine how the R179C variant influences chromatin accessibility. Explanted mouse aortic SMCs from WT and Acta2SMC-R179C/+ mice were plated at a density of 100,000 cells per 6 well plate, allowed to attach overnight, then serum starved for 24 hours. Two independently prepared wells from separate frozen vials of cells were used as replicates for the experiment. Cells were harvested, counted, then frozen and processed for ATAC sequencing (cell lysis, nuclear permeabilization, transposase reaction, indexed library amplification and sequencing using a commercially available kit (Active Motif, Carlsbad, CA).