Journal: Stem Cell Research & Therapy
Article Title: miR-181b/Notch2 overcome chemoresistance by regulating cancer stem cell-like properties in NSCLC
Figure Lengend Snippet: 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
Article Snippet: The slides were incubated with Notch2 (1:200 dilution, Abclonal), CD133 (1:200 dilution, JKSJ-orb372326), and SOX2 (1:100 dilution, Abcam-ab92494) primary antibodies, stained using DAB, and counterstained using haematoxylin.
Techniques: Expressing, Western Blot, Luciferase, Activity Assay, Mutagenesis, Transfection, Negative Control, Light Microscopy, Flow Cytometry, Cytometry, Real-time Polymerase Chain Reaction