Biotinylated (Dharmacon, Lafayette, CO, USA) pull-down assay was performed as described earlier [12 (link)]. Briefly, 1 × 106 TE7 cells were transfected with biotin-labelled miR-214-3p or control miR at a final concentration of 50 nM for 48 h. Whole cell lysates were incubated at 4 °C overnight with 50 μl/sample of streptavidin-Dyna beads (Invitrogen, Carlsbad, CA, USA) coated with yeast tRNA (Ambion, Austin, TX, USA). Next day, beads were washed thoroughly and RNA was isolated using TRIzol (Invitrogen) and standard chloroform-isopropanol method and then subjected to PCR as explained above.
Free full text: Click here
