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. 2011 Sep 12;6(9):e24584. doi: 10.1371/journal.pone.0024584

Figure 5. Decrease in CD15+ and CD133+ expression in Daoy cells under hypoxia condition, upon miR-34a overexpression.

Figure 5

A, B. Real-time PCR showing CD15 (A) and CD133 (B) expression in Daoy cells grown under normoxia and hypoxia conditions (as indicated) for 12 h, after 12 h of infection with AdV-miR-34a or AdV-GFP-mock viruses. Fold-changes are shown with respect to CD15 and CD133 expression, as measured in AdV-GFP-mock infected cells. Data are means ±standard deviations of 3 experiments, each carried out in triplicate *: p<0.05. Real-time PCR reactions were normalized to β-Actin. C. Representative FACS analysis for CD15+ and CD133+ subpopulations in Daoy cells grown under normoxia or hypoxia conditions for 12 h, after 24 h of infection with AdV-GFP-miR-34a or AdV-GFP-mock. D. Representative Western blot for Daoy cells and two primary human MB cell lines (SaV-MB1 and ViV-dMB) at 72 h of treatment with a SNALP carrying miR-34a or with a SNALP-scrambled, performed using anti-CD133, anti-CD15 and anti-β-Actin antibodies. E. Representative Western blot with a Daoy miR-34a stable clone and a Daoy empty-vector stable clone, using an antibodies panel against: Ak, Akt-S473, STAT3-S727, MEK1/2 S217-221, MARCK S152-156 and β-Actin. F. Representative Western blot of normal mouse cerebellum, and Patch+/- P53-/- and primary Patch+/- P53-/- mouse MB cell lines, at 48 h from infection with AdV-GFP-miR-34a or AdV-GFP-mock viruses, carried out using anti-Dll1 and anti-β-Actin antibodies.