FIGURE 3.

No evidence for critical epigenetic changes upon DHM treatment in keratinocytes. (A). Direct pyrosequencing after bisulfite conversion of the LINE-1 sequence covering methylation of 4 representative CpGs. Bar plot represents mean methylation difference of the analyzed LINE-1 sequence between 20 µM DHM and DMSO treatment in % (n = 4). (B) DNA damage assay. Bar plots depicting number of DNA double strand breaks per cell in primary human keratinocytes upon DHM treatment (blue) and positive control H₂O₂ (yellow) compared to the DMSO control (gray). *p < 0.05 (n = 6; 2-way ANOVA). (C) Apoptosis assay. Bar plots depicting caspase 3/7 activity in primary human keratinocytes upon DHM treatment (blue) and positive control staurosporine (orange) compared to the DMSO control (gray). *p < 0.01 (n = 10; one-way ANOVA, Dunnett’s multiple comparison test).