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. 2017 Feb 8;8(12):19866–19878. doi: 10.18632/oncotarget.15189

Figure 4. miR-214 mediated VSMCs gene expression through modulating QKI.

Figure 4

A-F. Over-expression of QKI up-regulates VSMCs gene expression, while knockdown of QKI down-regulates their expression. Day 2~3 differentiating ESCs were transfected with control (GFP) or QKI over-expression plasmid (GFP-QKI), or control siRNA (NC) or QKI specific siRNA (siRNA-QKI), respectively. Transfected cells were cultured in VSMCs differentiation medium for another 48 or 72 hours. Total RNAs and proteins were harvested and subjected to RT-qPCR and Western Blotting analyses, respectively. QKI and its isoforms were upregulated (A and B), resulting in VSMCs marker genes downregulation (D and F), while siRNA-QKI (C) increased marker genes expression (E and F) at both RNA and protein level. (G) Knock-down of QKI abolished the inhibitory effect of miR-214 inhibition on VSMCs gene expression. Day 2~3 differentiating ESCs were co-transfected with miR-214 inhibitor, QKI siRNA or respective negative control as indicated in the figure, and cultured for further 48-72 hours. Total proteins were harvested and subjected to Western blotting assays. The data presented here are representative or mean±S.E.M. of three independent experiments, *P<0.05, #P<0.05 (versus respective control)