Additional file 6 of Epigenetic reactivation of tumor suppressor genes with CRISPRa technologies as precision therapy for hepatocellular carcinoma

Additional file 6: Figure S5. Related to Fig. 5. Tiling the promoter of MT1M and TTC36 tumor suppressor genes with a CRISPRa toolbox in Hep3B HCC cells. A-B Fold change in MT1M (A) and TTC36 (B) mRNA expression evaluated by qRT-PCR 72 h after transient transfection in Hep3B cells. Cells were transfected with combinations of CRISPRa along with a mix of four gRNAs (MIX 4G), or with no gRNA (NO G) as control. Relative gene expression was normalized and compared to cells transfected with empty vector control (EV) for statistical analysis. Data presented as means ± SEM (n = 3), and P-values were determined by one-ANOVA with Dunnett's multiple comparisons test (****P < 0.0001). SpdCas9 Streptococcus pyogenes deactivated Cas9 protein adopted for epigenome engineering, VPR VP64, p65, Rta, MS2 RNA aptamer, MCP MS2-coat protein, HSF1 heat shock factor 1, TET1-CD Ten-Eleven Translocation methylcytosine dioxygenase 1-catalytic domain, MIX 4G combination of four gRNAs.