miR-17 and miR-20a contribute to Hbp1 regulation in NN10#5 and 7451#44 cells.

<p>A, B, C and D : Parental NN10#5 cells and their derivative #17-92a, #17-92b and #17-20 were cultured for two days in the presence or absence of Dox before Western blot and qRT-PCR analyses of known targets of miR-17-92 cluster. A: Western blot analysis of non-regulated targets in NN10#5 cells. B: Histograms show relative levels of hbp1 mRNA in the indicated Dox-treated cells normalized to that in untreated cells (means and standard deviations from 3 experiments). Asterisks indicate significant differences induced by Dox treatment (p<0.05). C: Western blot analysis of Hbp1 protein. D: Quantification of Hbp1 protein level variations in response to Dox between the indicated cells (+Dox/-Dox ratios of Hbp1 signal standardized to Actin signal determined by densitometric analysis of Western blot shown in C). E, F, G and H : 745#44 cells were grown as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046799#pone-0046799-g001" target="_blank">Figure 1</a> for two days in the presence or absence of HMBA or Dox as indicated. E: Western blot analysis of non-regulated targets. F: Histograms show relative levels of hbp1 mRNA (typical results from two different experiments). G: Western blot analysis of Hbp1 protein (typical results from two different experiments). H: Quantification of relative levels of Hbp1 protein standardized to actin.