Figure 3: Argonaute 1 and promoter-associated RNA coordinate CDH1 gene silencing. | Nature Communications

Figure 3: Argonaute 1 and promoter-associated RNA coordinate CDH1 gene silencing.

From: A promoter-proximal transcript targeted by genetic polymorphism controls E-cadherin silencing in human cancers

Figure 3

(a) CDH1 mRNA (left) and protein levels (right) in PC3 cells transfected for 72 h with siAGO1 or siGL3. (b) CDH1 mRNA in DU145, LNCaP and PrECs transfected with siAGO1 or siGL3 and collected after 72 h. (c) AGO1 binding to the CDH1 promoter determined by ChIP (primers F-281/R-139). (d) Binding of AGO1 to S-paRNA determined by RNA-ChIP. (e) Expression of S and AS-paRNA (top) and binding to AGO1 (bottom) determined by RIP in PC3 cells transfected with paRNA and AGO1 expression vectors. CD44 was used as positive control for AGO1-bound mRNA. (f) Binding of truncated S-paRNA transcripts to AGO1 determined by RIP. Arrows, forward and reverse primer sets for each transcript. (g) Histone mark modifications at the CDH1 promoter in PC3 cells transfected with si217 (top) or siAGO1 (bottom) and control siGL3. (h) Binding of SUV39H1 to the CDH1 promoter in PC3 cells transfected with si217 (left) or siAGO1 (right) and siGL3. Data are mean±s.d. of three independent determinations. Student’s t-test was used for P value assessment. *P<0.05.

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