Cultured hcSMCs were washed once with PBS and lysed directly with the lysis buffer in the flasks, and total RNA and DNA were extracted using RNA/DNA/Protein Purification Plus Kit (Norgen #47700). The DNA samples was quantified using Qubit® 2.0 Fluorometer (Life technologies). One hundred ng of genomic DNA spiked with one ng of unmethylated lambda DNA (Promega) was served for tPBAT protocol (Miura F et al, 2019).