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Fig. 3 | Clinical Epigenetics

Fig. 3

From: MAL expression downregulation through suppressive H3K27me3 marks at the promoter in HPV16-related cervical cancers is prognostically relevant and manifested by the interplay of novel MAL antisense long noncoding RNA AC103563.8, E7 oncoprotein and EZH2

Fig. 3

Knockdown of E7 by siRNA against E7 in HPV16-positive CaCx cell line, SiHa. A TaqMan-based qRT-PCR revealed 69% knockdown in HPV16 E7 expression in transfected group (T-SiHa) compared to the untransfected (UT-SiHa). B Upper panel: Western blot showing knockdown of E7 protein (19KDa) in transfected group (T-SiHa) compared to the untransfected (UT-SiHa). Lower panel: Western blot performed for house-keeping protein β-actin (42 KDa) for UT-SiHa (untransfected SiHa) and T-SiHa (transfected SiHa). The data are represented as mean ± SD from two independent experiments. Chromatin immunoprecipitation (ChIP)-qPCR with H3K27me3 antibody (C, D). C Fold enrichment showing the depletion of H3K27me3 at the MAL promoter in the transfected (T SiHa) when compared to that of untransfected (UT SiHa). EVX1 promoter is shown as the positive control for H3K27me3 in UT and T SiHa. The data are represented as mean ± SD from three independent experiments. D % input showing the depletion of H3K27me3 at the MAL promoter in transfected (T SiHa) when compared to that of untransfected (UT SiHa). Effect of E7 knockdown in HPV16-positive SiHa cells (E–G). E Relative expression of MAL F EZH2 G AC103563.8, respectively, in transfected (T SiHa) when compared to that of untransfected (UT SiHa). These plots depict the relative gene expression, i.e. expression of the gene—expression of the house-keeping gene (GAPDH). Relative gene expression is inversely proportional to the fold change. Thus, higher relative gene expression designates lower fold change. The data are represented as mean ± SD from two independent experiments

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Clinical Epigenetics

ISSN: 1868-7083

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