Figure 5. Hypoxia stimulates cancer stem cell associated phenotypes in a miR-200b dependent manner.
HMLER cells overexpressing miR-200b or GFP (n=3) grown for 7 days under 1% O2 or control conditions. (a,b) qRT-PCR analysis was performed to measure (a) mature miR200b levels, (b) DICER, ZEB1 and E-cadherin mRNA level where RPL13A served as control (n=3). (c) Representative FACS analysis using antibodies specific for CD44 and CD24. (d) Quantification of the percentage of cells with CD44highCD24low for 3 independent experiments. (e) Sphere formation assay, number spheres formed per 500 cells plotted (n=3). (f) Model of hypoxia mediated EMT and stemness. Error bars represent s.e.m. P values obtained with Student’s t-test. *P<0.05, **P<0.01, ***P<0.001