Figure 1
From: A novel RT-QPCR-based assay for the relative quantification of residue specific m6A RNA methylation
(A) Graphical representation of retrotranscription followed by PCR of HPRT (left). Agarose gel after PCR of the retrotranscribed samples using BstI and MRT enzymes (right). (B) Graphical representation of retrotranscription followed by PCR of TUG1 lncRNA. m6A mark (in green) was described by SCARLET method (Liu, N. et al.9) (left). Agarose gel after PCR of the retrotranscribed samples using m6A (+) and (−) primers and BstI and MRT enzymes. (C) m6A-RT-QPCR using different time points (5 min, 15 min and 30 min) for the retrotranscription of methylated TUG1 and unmethylated HPRT mRNAs. RNA was extracted from three different passages of HCT15 intestinal cell line. (D) Relative quantification of m6A levels of TUG1 and HPRT in Caco 2 intestinal cells. Data are represented as the mean and standard error of three independent experiments. **p-value < 0.01 based on paired Student’s t-test. (E) Comparison of m6A IP data retrieved from the Met-DB v2.0 database and relative m6A RT-QPCR results in different genes using the HEK293 cell line. Genes in bold showed concordant results using both methods. Grey boxes represent negative (non-methylated) and black boxes represent positive (methylated) results. Relative RT-QPCR data correspond to the mean and standard error of four independent experiments.
