Taqman universal master mix 2 non ung kit

The qMSP analysis was performed following a previous protocol 40 (link). Briefly, the DNA methylation inhibitor AZA (5-Aza-2'-deoxycytidine)-treated cells were subjected to DNA isolation using a PureLink^TM Genomic DNA Mini Kit (Thermo Fisher Scientific, #K182001) according to the manufacturer's protocol. After quantification with a Nanodrop spectrophotometer, 1 µg of genomic DNA from each sample was treated with sodium bisulfite using the EZ DNA Methylation-GOLD Kit (Zymo Research, Tustin, CA, USA, #D5006) following a protocol provided by the manufacturer. The resulting genomic DNA was used for qMSP analysis using the TaqMan^® Universal Master Mix II non-UNG Kit (Thermo Fisher Scientific, #4440038) with primers listed in Supplementary Table-4.

The qMSP assay was carried out as described previously 43 (link). Genomic DNA was isolated from tissues or cells using a Monarch Genomic DNA Purification Kit (New England Biotechnology, Beijing, China, #T3010L) following the manufacturer's protocol. A total of 2.0 μg genomic DNA from each sample was treated with an EZ DNA Methylation-GOLD Kit (Zymo Research, Tustin, CA, USA, #D5006) in which sodium bisulfite converted unmethylated cytosines but not methylated cytosines to uracil. The modified DNA was subjected to qMSP analysis using a TaqMan^® Universal Master Mix II non-UNG Kit (ThermoFisher Scientific, #4440038) with primers included in Supplementary Table-5.