Fig. 3: SP1 regulated the methylation of EphA7 by interfering with DNMT1. | Cell Death & Disease

Fig. 3: SP1 regulated the methylation of EphA7 by interfering with DNMT1.

From: Targeted demethylation of the EphA7 promoter inhibits tumorigenesis via the SP1/DNMT1 and PI3K/AKT axes and improves the response to multiple therapies in cervical cancer

Fig. 3

A The JASPAR (http://jaspar.genereg.net/), CONTA V3 (http://bioit2.irc.ugent. be/contra/v3), and hTFtatget (http://bioinfo.life.hust.edu.cn/ hTFtarget) tools were integrated to predict transcription factor binding in the promoter of EphA7. SP1 and MAZ were found to directly interact with EphA7 on the basis of the Pathway Commons (https://www.pathwaycommons.org). B The mRNA and protein expression analysis showed that total SP1 and MAZ were significantly expressed in SiHa cells. C The in situ pull-down results verified that SP1 and MAZ were located in the promoter region of EphA7 in SiHa cells. D Pathway commons tools indicating that SP1 and MAZ directly interact with EphA7 and DNMT1. Promoter methylation of EphA7 was associated with DNMT1 (P < 0.05) in TCGA-CESCs (n = 306). E Western blotting was performed to demonstrate that SP1 was silenced in SiHa-SP1 knockout cells (SiHa-SP1-KO) and that the protein expression levels of EphA7 were increased in SP1-KO cells (n = 3). F The mRNA expression levels of EphA7 were increased, and the expression of DNMT1 was decreased in SP1-KO cells (n = 3). G Knockout of SP1 markedly reduced the methylation level of EphA7 in SiHa cells. H, I The results of rescue assays showed that SP1 overexpression reversed the changes in the mRNA (P < 0.05) and protein expression of EphA7 in the SP1-KO group. Error bars represent the means ± SDs, P values were calculated using two-tailed unpaired Student’s t tests (E–G) and Spearman’s analysis (D). *P < 0.05, **P < 0.01,  ***P < 0.001.

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