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Fig. 3 | Stem Cell Research & Therapy

Fig. 3

From: miR-181b/Notch2 overcome chemoresistance by regulating cancer stem cell-like properties in NSCLC

Fig. 3

miR-181b suppresses Notch2 signalling to inhibit CSC traits. a Venn diagrams show the number of genes identified as potential targets of miR-181b in three predictive programs: TargetScan, miRanda, and miRBD. Notch2 expression in adherent A549/DDP and A549 cells or cell spheres was tested by Western blotting. b Relative luciferase activity was evaluated after wild-type or mutant 3′-UTR reporter plasmids were co-transfected with pGLE-miR-181b or miR-181b-NC in H1299 cells. c A549 cells were transfected with miR-181b inhibitors or the control and then treated with si-Notch2 or negative control. Notch2 expression was analysed by Western blotting. d The number of tumourspheres was counted, and the morphology was observed under a light microscope. e The IC50 value was determined by CCK assay with different concentrations of cisplatin. The apoptotic percentage (f) and CD133+ cells (g) were evaluated by flow cytometry. h The mRNA levels of KLF4, SOX2, NANOG, CD133, and ALDH were measured by qPCR. i Western blotting showed Notch2, NICD2, HES1, and HEY1 expression after A549/DDP and A549 cells were transfected with miR-181b mimics, miR-181b inhibitors, or the control. Bars represent 200 μm for low-power lens and 50 μm for high-power lens. Data are presented as the mean ± SD. *p < 0.05; **p < 0.01; ***p < 0.001

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