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. 2016 Jun 24;6:28436. doi: 10.1038/srep28436

Figure 6. Snail mediates repression of miR-128 in response to Aurka overexpression.

Figure 6

(A) AurkA overexpression does not promote Snail protein increase as found in MCF-AurkA (AurkA+) versus control cells (empty) Evaluation of Snail protein levels in MCF-7 and MCF-AurkA. In contrast, AurkA knock-down severely impaired Snail protein levels in MDA-shAurkA (sh5, sh7, sh8) and KBr2-shAurkA (shAurkA) in comparison with control cells (empty). (B) AurkA antibody pulls down Snail in MCF7 and MDA-MB-231 cells after a co-immunoprecipitation assay (IP anti-AurkA). As control, protein lysates from both cell lines were tested for Snail protein levels (control). A marker was loaded in the last lane as a control of molecular weight. (C) AurkA overexpression in MCF-7 cells induced nuclear translocation of Snail (MCF, AurkA+). Control cells show a moderate cytoplasmic staining (MCF, empty). Nuclear accumulation of Snail in MDA-MB-231 cells (MDA, empty) was inhibited by AurkA knock-down (MDA, shAurkA). Nuclei were counterstained with DAPI. Collectively our findings supports a signaling pathway showing that AurkA may promote nuclear translocation of Snail to repress miR-128 gene transcription. As results, wnt3a mRNA accumulates increasing Wnt3a protein levels. (D) On the left, Q-PCR confirmed association between AurkA and miR-128 levels in 32 human tissues from breast cancer patients. Asterisk highlights samples were association is missing. On the right, Graphic representation of of samples (percentage, %) showing a strong correlation AurkA-mir-128 (AurkA > 1.5/miR-128 < 1.5 and AurkA < 1.5/miR-128 > 1.5), or where AurkA-miR-128 correlation is missing (AurkA > 1.5/miR-128 > 1.5 and AurkA < 1.5/miR-128 < 1.5).