Fig. 4: Validation of ASB2 as a direct target of miR-29c. | Cell Death & Disease

Fig. 4: Validation of ASB2 as a direct target of miR-29c.

From: High-throughput analysis of the RNA-induced silencing complex in myotonic dystrophy type 1 patients identifies the dysregulation of miR-29c and its target ASB2

Fig. 4

a Undifferentiated myogenic cells were transfected with miR-29c- or scramble (SCR)-expression vectors and the levels of ASB2 and CDC42 were analyzed by western blotting. A representative blot is shown. Vinculin (VCN) is shown as loading control. The histogram shows the quantitation of the expression levels of ASB2 and CDC42 normalized to vinculin in cells expressing miR-29c vs cells expressing scramble, referred as 1 (n = 3; *P < 0.05; ***P < 0.001). b HEK-293 cells were transfected with empty Firefly luciferase reporter vector (pMIR-REPORT) or derived constructs containing either an intact miR-29c-binding site (wt), or a mutated miR-29c-binding site (mut) in ASB2 3′UTR target region. Each plasmid was co-transfected with a plasmid encoding renilla luciferase along with miR-29c- or scramble (SCR)-expression vectors. Activities of each pMIR-REPORT construct were normalized first to Renilla luciferase and then to empty vector activities. Luciferase (luc) activity in cells transfected with miR-29c is shown relative to activity in cells transfected with scramble RNA, referred as 1 (n ≥ 11; **P < 0.01)

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