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. 2013 Aug 28;41(21):9688–9704. doi: 10.1093/nar/gkt680

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© The Author(s) 2013. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 10. — miR-17 represses TIMP3, PTEN and p21 expression in tumor sections. (a) Cell lysate prepared from miR-17 or control-transfected cells was analyzed on western blot for p21 and PTEN expression. Repression of p21 and PTEN expression was seen in cells transfected with miR-17. Staining for GAPDH from the same membrane confirmed equal loading. (b) Lysate prepared from tumors formed by the miR-17 or control-transfected cells was also analyzed for p21 and PTEN expression. Decreased p21 and PTEN levels were seen in the miR-17 tumors compared with the control. (c) Sections from the miR-17 and control tumors were probed with anti-PTEN and anti-p21 antibodies. In general, tumor sections from the miR-17 transfected cells exhibited much lower levels of PTEN and p21 than the control. (d) Individual and combined effects of miR-17-5p mimic (17-5pM) and miR-17-3p mimic (17-3pM) on p21 and PTEN expression was tested. 17-3pM minimally repressed p21 and PTEN expression, but 17-5pM showed a clear effect in repressing p21 and PTEN expression (left). DU145 cells transfected with a control (middle) or miR-17 vector (right) were treated with miRNA inhibitors. The miR-17-5p inhibitor (17-5pI) but not 17-3pI enhanced p21 and PTEN expression.