Figure 6.

Knockdown of NF90 decreases pri-miR-590 and increases mature miR-590-5p. (a) whole-cell lysates were prepared from SW620 cells transfected with the indicated siRNAs and analyzed by immunoblotting with anti-NF90 and anti-α-tubulin antibodies. (b) The amount of pri-miR-590 was analyzed using qRT-PCR. GAPDH functioned as an internal control. (c) RNAs were isolated from SW620 cells transfected with the indicated siRNAs and analyzed for the amount of mature miR-590-5p. U6 was used as an internal control. (d) Whole-cell lysates were prepared from SW480 cells and analyzed by immunoblotting with anti-NF90 and anti-α-tubulin antibodies. (e and f) The amount of pri-miR-590 and mature miR-590-5p were analyzed by qRT-PCR. (g) NF90 binds to pri-miR-590 in vitro. EMSA was performed with the pri-miR-590 probe and without (−) or with recombinant NF90 protein alone at the indicated protein concentrations. *P<0.05, **P<0.01, and ***P<0.0001