Fig. 6.

Modulation of ZAR1 promoter methylation offers therapeutic approach to ZAR1 reactivation. a Hypermethylation of ZAR1 by DNMT3A using ZAR1 promoter (pRLnull) in HEK and HeLa for indicated time points followed by CoBRA methylation analysis and b according quantification by pyrosequencing. c Pharmacological DNMT inhibition by 0.5 μM/1 μM 5-aza-2′-deoxycytidine (Aza) together with ZAR1 targeted VP160 overexpression activates ZAR1 expression. d ZAR1 endogenous reexpression by epigenetic editing through overexpression of ZAR1 guided p300, VP160, and TET1 in HeLa, HEK, and HCT116. e Promoter methylation of HeLa, HEK, and HCT116 by CoBRA and f quantified by pyrosequencing. g ZAR1 endogenous reexpression by epigenetic editing through ZAR1-guided TET1 upon overexpression h is guide-combination dependent. i, j ZAR1 reexpression by overexpressed TET1 (guided by ZAR1 oligos) is accompanied by ZAR1 demethylation by pyrosequencing