Treatment with complementary DNA-LNA chimeras knocked down endogenous miRNAs in MDA-MB-231 TNBC cells.

<p><b>A</b>: qPCR of miR-17-5p 12 hr and 48 hr post-transfection with anti-miR-17-5p. <b>B</b>: qPCR of miR-21-5p 12 hr and 48 hr post-transfection with anti-miR-21-5p. Results represent absolute values of miRNA/internal control U6 normalized to mock transfected. Values are the average of three measurements ±.</p

Predicted anti-miR-17-3p binding sites in the 3'UTR of <i>PDCD4</i> and <i>PTEN</i> mRNAs as a mimic of miR-17-5p passenger strand.

<p>^aThe potential oncomiR:mRNA binding sites were identified by rna22, Targetscan, or miRanda. ^bTop strand is mRNA (5' ➔ 3') and the bottom strand is oncomiR (3' ➔ 5'). ^cCalculated at <a href="http://mfold.rna.albany.edu/?q=mfold" target="_blank">http://mfold.rna.albany.edu/?q=mfold</a>.</p

Schematic view of competition between anti-miR-17-5p and miR-17-5p for inhibition of <i>PDCD4</i> mRNA.

<p>Schematic view of competition between anti-miR-17-5p and miR-17-5p for inhibition of <i>PDCD4</i> mRNA.</p

Both miR-17-5p and miR-17-3p can directly modulate the translation of <i>PDCD4</i> and <i>PTEN</i>.

<p>A: PDCD4 and PTEN protein Western blots at 48 hr post transfection with miR-17-5p mimic or miR-17-3p mimic. β-actin was used as loading control. Values are the average of three blots ± s.d. after normalization to β-actin and to control/treatment group. Each blot was subjected to gamma setting adjustments. B—E: Luciferase activity after co-transfecting MDA-MB-231 cells with indicated luciferase reporter constructs in the presence or absence of miRNA mimic. All luciferase signals from pMir-report firefly luciferase vectors are normalized to signals from pRL-TK Renilla luciferase vector. The ratio of normalized signal in the presence of mimic to signal in the absence of mimic for each construct is then calculated. The pMir-report luciferase vector is used as negative control. Results represent fold changes of the above ratio relative to vector control. Values are the average of at least three measurements ± s.e.m * indicates p<0.05, ** indicates p<0.01.</p

Predicted miR-17-3p passenger strand binding sites in the 3'UTR of <i>PDCD4</i> mRNA.

<p>^aThe potential oncomiR:mRNA binding sites were identified by rna22, Targetscan, or miRanda. ^bTop strand is mRNA (5' ➔ 3') and the bottom strand is oncomiR (3' ➔ 5'). ^cCalculated at <a href="http://mfold.rna.albany.edu/?q=mfold" target="_blank">http://mfold.rna.albany.edu/?q=mfold</a>.</p

Predicted anti-miR-17-5p binding sites in the 3'UTR of <i>PTEN</i> mRNA as a mimic of miR-17-3p passenger strand.

<p>^aThe potential oncomiR:mRNA binding sites were identified by rna22, Targetscan, or miRanda. ^bTop strand is mRNA (5' ➔ 3') and the bottom strand is oncomiR (3' ➔ 5'). ^cCalculated at <a href="http://mfold.rna.albany.edu/?q=mfold" target="_blank">http://mfold.rna.albany.edu/?q=mfold</a>.</p

Predicted miR-21-5p guide strand binding sites in the 3'UTR of <i>PDCD4</i> mRNA.

<p>^aThe potential oncomiR:mRNA binding sites were identified by rna22, Targetscan, or miRanda. ^bTop strand is mRNA (5' ➔ 3') and the bottom strand is oncomiR (3' ➔ 5'). ^cCalculated at <a href="http://mfold.rna.albany.edu/?q=mfold" target="_blank">http://mfold.rna.albany.edu/?q=mfold</a>.</p

Predicted anti-miR-17-5p binding sites in the 3'UTR of <i>PDCD4</i> mRNA as a mimic of miR-17-3p passenger strand.

<p>^aThe potential oncomiR:mRNA binding sites were identified by rna22, Targetscan, or miRanda. ^bTop strand is mRNA (5' ➔ 3') and the bottom strand is oncomiR (3' ➔ 5'). ^cCalculated at <a href="http://mfold.rna.albany.edu/?q=mfold" target="_blank">http://mfold.rna.albany.edu/?q=mfold</a>.</p

Effect of complementary DNA-LNA chimeras on the <i>PDCD4</i> and <i>PTEN</i> mRNA levels in MDA-MB-231 cells.

<p><b>A</b>: qPCR of <i>PDCD4</i> mRNA at 12 hr and 48 hr after transfection. <b>B</b>: qPCR of <i>PTEN</i> mRNA at 12 hr and 48 hr after transfection. <b>C</b>: Relative expression of <i>PDCD4</i> mRNA from 12 hr to 48 hr after transfection with anti-miR-17-5p and anti-miR-17-3p. <b>D</b>: Relative expression of <i>PTEN</i> mRNA from 12 hr to 48 hr after transfection with anti-miR-17-5p and anti-miR-17-3p. Results represent absolute values of miRNA/internal control gene <i>GAPDH</i> normalized to mock transfected. Values are the average of three measurements ± s.d. * indicates p<0.05, ** indicates p<0.01.</p

Sequence alignment of the EGFR ligands.

<p>a) Shown are the seven ligands used in the computational studies. Sequences of only the EGF like domains of each ligand were used in the alignment. “*” represent 100% conservation while “.” and “:” represent partial sequence conservation.</p