Fig. 4

Experimental validation of the target genes of the miR-130b using dual luciferase assay. a The luciferase reporter assay in cumulus cells co-transfected with miR-130b precursor or inhibitor and the pmirGLO vector construct harboring of the 3′ UTRs of the SMAD5 gene (SMAD5GLO) or non-target sequence (Non-targetGLO). b The luciferase reporter assay in cumulus cells co-transfected with miR-130b precursor or inhibitor and the pmirGLO vector construct harboring of the 3′ UTRs of the MSK1 gene (MSK1GLO) or non-target sequence (Non-targetGLO). The firefly and renilla activity ratio in cells transfected only with vector construct harboring the 3′ UTR of the miR-130b target genes or cells co-transfected along with the miRNA scramble sequence or the firefly and renilla activity ratio of cells co-transfected with miR-130b precursor or inhibitor along with the vector construct harboring the non-target sequence of miR-130b were used as controls. The assay was performed 24 h post transfection. Bars bars with different letters are statistically significant (p < 0.05). RE; relative expression; FL; firefly luminescent; RL; renilla luminescent. Bars graphs represent the mean ± standard deviation (SD) and the experiment was repeated three times in three independent samples