Fig. 4

The expression of differentially methylated genes and genes encoding endothelial cell adhesion markers (i.e., ICAM1, VCAM1, and SELE) in site-specific methylation model HUVECs (dCase9-Dnmt3a) and negative control HUVECs. (a) TRIM6 model: (left) methylation level at cg15137954 is higher in dCas9-Dnmt3a engineered cells than in negative control cells; (center) higher methylation is associated with decreased TRIM6 expression; (right) decreased expression of VCAM1 and SELE can be observed in dCas9-Dnmt3a engineered cells. (b) FLRT2 model: (left) methylation level at cg16800165 is higher in dCas9-Dnmt3a engineered cells than in negative control cells; (center) FLRT2 expression is significantly decreased in dCas9-Dnmt3a engineered cells; (right) expression of genes encoding endothelial cell adhesion markers is significantly increased in dCas9-Dnmt3a cells. (c) TLN2 model: (left) methylation at cg15949239 is higher in dCas9-Dnmt3a cells than in the negative control cells; (center) TLN2 expression is significantly increased in dCas9-Dnmt3a cells; (right) expression of genes encoding endothelial cell adhesion markers is significantly decreased in dCas9-Dnmt3a engineered cells when compared to negative control cells. Relative mRNA expression of dCase9-Dnmt3a to negative control was calculated using the 2(-Delta Delta C(T)) method and normalized with the reference gene Actin (3 repeated measurements per group). Data are mean ± SEM (standard error of mean). A Student t test was used to compare the group difference in gene expression levels. *P < 0.05; **P < 0.01; ***P < 0.001