Figure 4.

(a) The assessment of targeted editing using the All-in-one system in CM150-post. The graph illustrates the average DNA methylation levels for both the target region and the entire amplicon of EBF3 in CM150-post. It compares methylation levels between unedited and edited samples, represented in absolute values. Additionally, it presents the relative change in DNA methylation levels between these two samples (denoted as ∆). Following this, heatmaps are used to depict the methylation status for each CpG site within the target region, with unmethylated sites in blue, methylated sites in red, and unaligned sites in white. These heatmaps were generated based on 500 randomly selected sequencing reads. (b) The assessment of targeted editing using the All-in-one system in WM266-4. Similar to CM150-post, the WM266-4 graph displays the average DNA methylation levels for target and entire amplicon region of EBF3 with corresponding heatmaps. Additionally, the error bars in the graph indicate the mean value with its standard error (SEM) calculated from three separate biological replicates. Statistical analysis using a t-test yielded p-values > 0.05 for both cell lines (**** p < 0.0001). (c) The Bland–Altman (BA) plot illustrates a comparison between the means of CpG methylation for both unedited CM150-post and WM266-4 cell lines. This analysis involved two technical replicates for each cell line, as depicted in the figure. The proximity of each data point to the x-axis indicates the level of similarity between the means of the two replicates, representing the degree of difference. On the other hand, the horizontal position of each data point on the x-axis corresponds to the average of the two means. The two dashed lines on the plot mark the upper and lower 95% limits of agreement, which are −0.01328 to 0.005074 for CM150-post and −0.02936 to 0.020790 for WM266-4, respectively.