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. 2016 Jun 1;7(29):45134–45143. doi: 10.18632/oncotarget.9761

Figure 5. EAF2 is an EZH2 target gene involved in the regulation of HIF1α.

Figure 5

(A) IHC analysis of EZH2 and EAF2 expression in glioblastoma specimens and samples of normal brain tissue (400×). (B) Correlation between expression of EZH2 and EAF2 in 105 blioblastoma specimens (P < 0.05). (C) Levels of EAF2 and HIF1α protein were measured by immunoblotting U251 cells transfected with control vector, EZH2-shRNA or pJAX-EZH2. β-actin was used as a loading control. (D) EAF2 expression was silenced or not in U251 cells transfected with SC-shRNA or EZH2-shRNA. Immunoblots showing EAF2 and HIF1α levels in the indicated cells. β-actin was used as a loading control. (E) Immunoblots showing H3K27me3 and H3K9me3 levels in control and EZH2-overexpressing U251 cells. β-actin was used as a loading control. (F) U251 and T98G cells were subjected to ChIP analysis using antibodies against H3K27me3 or EZH2 with primers targeted to the promoter region of EAF2. Isotype matched IgG was used as a negative control. Data are presented as the mean ± s.d. from three independent experiments. *P < 0.05, **P < 0.01.