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. 2010 Oct 29;5(10):e13764. doi: 10.1371/journal.pone.0013764

Figure 2. miR-31 negatively regulates WNT signaling pathway antagonists in normal respiratory epithelia and lung cancer cells.

Figure 2

A) miR-31 was over-expressed in SAEC, HBEC, Calu-6, and H841 cells via transient transfection of primary miR-31 constructs. qRT-PCR analysis confirmed high level miR-31 expression in miR-31-transfected relative to control cells. B) qRT-PCR analysis of Dkk-1 and DACT3 expression levels in SAEC, HBEC, Calu-6, and H841 cells with or without over-expression of miR-31. Over-expression of miR-31 decreased Dkk-1 and DACT3 in all four cell lines. C) qRT-PCR analysis demonstrating decreased levels of endogenous miR-31 in SAEC, HBEC, Calu-6, and H841 cells following transient transfection of antisense-miR-31(Zip-miR-31) constructs relative to controls. D) qRT-PCR analysis of Dkk-1 and DACT3 expression levels in SAEC, HBEC, Calu-6 and H841 cells with or without down-regulation of miR-31. Knock-down of miR-31 enhances basal levels of Dkk-1 and DACT3 in these cells. E) qRT-PCR analysis demonstrating that knockdown of miR-31 partially blocks CSC-mediated decreases of Dkk-1 and DACT3 in SAEC. F) Western blot analysis of Dkk-1 and DACT3 expression in parental and vector control SAEC and Calu-6 cells, as well as SAEC and Calu-6 cells exhibiting constitutive over-expression or knock-down of miR-31. Densitometry values are normalized to b-actin control. Dkk-1 and DACT3 level were decreased, or somewhat enhanced in these cells following over-expression, or knock-down of miR-31, respectively.