Additional file 12: Fig. S3. Epigenetic modulation of RSPO2 at its promoter region does not affect the methylation status of intragenic CpG sites. A Schema illustrating the genomic position of the RSPO2 gene, the CpG sites analysed in the 450 K methylation platform, the gRNAs designed to modulate the DNA methylation status of its promoter region and the amplicons used for the pyrosequencing assays in the context of the modulated region (Amplicon pyrosequencing A, related to Fig. 5), or a region located downstream of this dCas9-targeted region (Amplicon pyrosequencing B, this figure). B Boxplot representing the DNA methylation levels of the indicated significant CpG sites located within the RSPO2 promoter region in CRC samples as determined by the 450 K array platform. Samples are coloured according to their dataset of origin, as indicated in Fig. 1A. C Barplots depicting the percentage of DNA methylation observed for the CpG sites included in a location downstream of the modulated differentially methylated region (Amplicon pyrosequencing B) in DLD1 and HCT116 cells in the context of control gRNA (grey) or gRNAs targeting this DMR (blue) in cells transduced with dCas9-TET1- or dCas9-TET1-IM-related chimeras. Data represent mean ± standard deviation of at least 3 independent experiments, and two-sided Welch’s t tests were applied for the different statistical comparisons versus each corresponding control condition. *p value < 0.05; n.s.—nonsignificant.