Fig. 2

TERT promoter DNA methylation in Hep-G2 after ASEE targeting the C228T mutated and wildtype allele. Average allele DNA methylation after ASEE with sgRNAs targeting the C228T position in Hep-G2 (A) sgRNA C228T-mut, on-target mutated allele is shown in purple; (B) off-target allele shown in purple; (C) sgRNA C228T-wt, on-target wildtype allele shown in blue; (D) off-target mutated allele shown in blue. Untreated Hep-G2 cells (UT) are shown in yellow and Hep-G2 transfected with scrambled sgRNA are shown in light green. The x-axis shows the position of CpG sites in the BS1 assay according to the reference sequence used in the analysis. A, B The purple line at position 123 shows the binding site for sgRNA C228T-mut. C, D The blue line at position 123 shows the binding site for sgRNA C228T-wt. Error bars are calculated based on at least three independent experiments. Black bar plots show the 1-q value after T Test comparison between Hep-G2 transfected with C228T-mut (A, B) or C228T-wt (C, D) and Hep-G2 transfected with scrambled sgRNA. q values > 1 were set as 1 for visualization purposes. Highly significant CpG sites are indicated by stars (q < 0.05)