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. 2013 Apr 26;8(4):e63032. doi: 10.1371/journal.pone.0063032

Figure 8. MiR-15a inhibits EWS clonogenic growth.

Figure 8

(A) Dual Renilla/Luciferase assay performed in Sk-ES-1 and RD-ES cells transfected with 50 nM LNA-anti-miR-15a or a negative control LNA, and the psiCHECK2 dual luciferase reporter with a miR-15a complementary binding site, or non-targeting binding site, in the 3′ UTR. Results represent the mean and SEM of two independent experiments, each performed in triplicate. “Basal” reporter activity corresponds to cells transfected with non-targeting reporter and negative control (non-targeting) LNA. (B) Clonogenic assay in Sk-ES-1 and RD-ES cell lines treated with 50 nM of a negative control LNA or an LNA targeting miR-15a. (C) Relative overexpression of miR-15a in Sk-ES-1 and RD-ES cells treated with 25 nM of a negative control mimic or miR-15a mimic was determined by qRT-PCR. miR-15a levels were normalized to an endogenous U6 control. Results represent the mean and SEM of three independent experiments, each performed in triplicate. (D) Clonogenic assay in Sk-ES-1 and RD-ES cell lines treated with 25 nM of a negative control mimic or a miR-15a mimic. Statistical significance was determined based on an unpaired student’s t-test comparison between the indicated treatments.