Quantitative analysis of differentially methylated regions was performed by BioMiao Biological Technology (Beijing, China) with the Agena MassARRAY platform (Agena, San Diego, USA). Briefly, genomic DNA was isolated from HL-7702 cell lines, and 2 μg DNA was treated with sodium bisulfite using an EZ DNA Methylation-Gold kit (ZYMO Research) according to the manufacturer’s instructions. The specific primers based on the reverse complementary strands of the
TFEB promoter were designed using EpiDesigner software (Agena), and the quantitative results for each CpG or multiple CpGs were analyzed with EpiTYPER
TM (Agena). The primer sequences were 5′-aggaagagagAGGTATTTAAGGGTATTTTTGGTGG-3′ and 3′-cagtaatacgactcactatagggagaaggctCCTATAATCCCAACATTTTAAAAAACC-5′. Bisulfite-modified DNA PCR amplifications were performed with a precycling hold at 94°C for 4 min and subjected to 45 cycles of 94°C for 20 s, 56°C for 20 s,72°C for 1 min, and a final extension at 72°C for 3 min. Further experimental analysis of the contents of DNA methylation was determined, as described previously
[18] (link).
TFEB promoter were designed using EpiDesigner software (Agena), and the quantitative results for each CpG or multiple CpGs were analyzed with EpiTYPER
TM (Agena). The primer sequences were 5′-aggaagagagAGGTATTTAAGGGTATTTTTGGTGG-3′ and 3′-cagtaatacgactcactatagggagaaggctCCTATAATCCCAACATTTTAAAAAACC-5′. Bisulfite-modified DNA PCR amplifications were performed with a precycling hold at 94°C for 4 min and subjected to 45 cycles of 94°C for 20 s, 56°C for 20 s,72°C for 1 min, and a final extension at 72°C for 3 min. Further experimental analysis of the contents of DNA methylation was determined, as described previously
[18] (link).
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