To perform real-time quantitative PCR with a LightCycler 480 (Roche Diagnostics, Basel, Switzerland), TaqMan Gene Expression Assays (Life Technologies) were used as a probe/primer set specified for the FAM119A (METTL21A) gene and CREB1 gene and a probe/primer set for the ACTB gene, a house-keeping gene that encodes β-actin. The expression level of the FAM119A (METTL21A) gene or CREB1 gene was normalized to the expression level of the ACTB gene for each sample, and relative mRNA expression levels were compared between the genotype subgroups for each gene. The experiments were performed in triplicate (separate experiments) for each sample, and average values were calculated for normalized expression levels.
