Figure 4: IsomiR-4534 binds to Argonaute 1 and controls CDH1 expression. | Nature Communications

Figure 4: IsomiR-4534 binds to Argonaute 1 and controls CDH1 expression.

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

Figure 4

(a) Protocol for miRNA identification by RIP, biotin-labelled RNA pull-down and small RNA sequencing. (b) Sequence of miR-4534 and isomiR-4534. Red, nucleotide changed in isomiR-4534. (c) miR-4534 and isomiR-4534 expression in prostate normal (PrECs), tumorigenic (ESE3kd-PrECs) and cancer cell lines (LNCaP, DU145 and PC3). (d) Nuclear and cytoplasmic distribution of miR-4534 and isomiR-4534 in PC3 cells. (e) miR-4534 and isomiR-4534 expression in DU145 cells 24 h after transfection with pre-miR-4534 (pMIR4534) or control expression vector. (f) Expression of miR-4534, isomiR-4534, miR-21 and miR-22 in PC3 cells after transfection of pri-miR-4534 targeting (siprimiR) or control (siGL3) siRNA. (g) miR-4534 and isomiR-4534 expression in DU145 cells transfected with pre-miR-4534 targeting gRNA constructs (del91-55 and del88-52) or empty vector and selected with puromycin. (h) CDH1 expression in PC3 (left) and LNCaP (right) cells after transfection with miR4534 and isomiR-4534 mimics or negative control (ctrl neg) miRNA. (i) CDH1 mRNA (top) and protein (bottom) expression in DU145 cells transfected with pri-miR-4534 targeting siRNA (siprimiR) or 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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