trans 4  (Chem Impex International)

Journal: Stem cell research

Article Title: Constitutive activation of Notch2 signalling confers chemoresistance to neural stem cells via transactivation of fibroblast growth factor receptor-1.

doi: 10.1016/j.scr.2019.101390

Figure Lengend Snippet: Fig. 1. High endogenous level of N2ICD in human glioma cells correlates with increased resistance to apoptosis. (A) Immunoblot detection of endogenous N2ICD protein in LN18 and LN229. (B) Immunoblot analysis of cleaved caspase-3 and cleaved PARP in LN18 and LN229 after treatment with etoposide for the indicated length of time. (C) Immunoblot ana- lysis showing N2ICD-knockingdown (siN2) in- creased etoposide-induced caspase-3 and PARP cleavage in LN229 glioma cells compared to LN229 cells transfected with non-specific control siRNAs (siCtrl). Numbers underneath cleaved caspase-3 and cleaved PARP represent relative band quantifica- tion. (D) Immunoblot analyses of caspase-3 and PARP cleavage after etoposide treatment of LN18 cells transfected with empty vector (Ø) or Notch2 plamid (N2).

Article Snippet: Hemmings N/A Experimental Models: Organisms/Strains Mouse: Myc-N2ICD/Nestin-Cre Tchorz et al. 2012 N/A Oligonucleotides Notch2 siRNA pools Santa Cruz Biotech sc-40,135 Primers for N2ICD, Hes-1, Bcl2-1 l and Mcl-1, see supplementary methods Recombinant DNA Software and Algorithms ImageJ, FIJI NIH https://imagej.nih. gov/ij/;https://fiji. sc/ Adobe Photoshop CS5.1 Adobe N/A Mendeley Desktop Elsevier www.mendeley. com GraphPad Prism 5 GraphPad N/A Other

Techniques: Western Blot, Lysis, Transfection, Control, Plasmid Preparation

Journal: Stem cell research

Article Title: Constitutive activation of Notch2 signalling confers chemoresistance to neural stem cells via transactivation of fibroblast growth factor receptor-1.

doi: 10.1016/j.scr.2019.101390

Figure Lengend Snippet: Fig. 4. Notch2 and FGF signalling crosstalk in NSCs. (A) Immunoblot analysis of the effect of GF removal (-GFs) on caspase-3 and PARP cleavage in N2 NSCs. (B) Immunoblot analysis of the effect of FGF2, EGF and insulin individual re-addition on caspase-3 and PARP cleavage. (C) Immunoblot showing the effect of selective removal of the GFs on caspase-3 and PARP cleavage. (D) Immunoblot of FGFR1 phos- phorylation levels in N2 NSCs compared to control NSCs (Ctrl). (E) Immunoblot showing the effect of FGFR specific inhibitor PD173074 on caspase-3 and PARP cleavage in N2 NSCs.

Article Snippet: Hemmings N/A Experimental Models: Organisms/Strains Mouse: Myc-N2ICD/Nestin-Cre Tchorz et al. 2012 N/A Oligonucleotides Notch2 siRNA pools Santa Cruz Biotech sc-40,135 Primers for N2ICD, Hes-1, Bcl2-1 l and Mcl-1, see supplementary methods Recombinant DNA Software and Algorithms ImageJ, FIJI NIH https://imagej.nih. gov/ij/;https://fiji. sc/ Adobe Photoshop CS5.1 Adobe N/A Mendeley Desktop Elsevier www.mendeley. com GraphPad Prism 5 GraphPad N/A Other

Techniques: Western Blot, Control

Journal: Stem Cell Research & Therapy

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

doi: 10.1186/s13287-018-1072-1

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 < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: We assessed the expression of the CD133 surface marker after transfection with miR-181b mimics (Invitrogen) or Notch2 siRNA (Santa Cruz Biotechnology).

Techniques: Expressing, Western Blot, Luciferase, Activity Assay, Mutagenesis, Transfection, Control, Negative Control, Light Microscopy, Flow Cytometry

Journal: Stem Cell Research & Therapy

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

doi: 10.1186/s13287-018-1072-1

Figure Lengend Snippet: Overexpression of miR-181b inhibits CSC characteristics in vivo. Mice were treated with different doses of A549/DDP sphere cells transfected with miR-181b agomir or miR-181b-NC agomir. Representative images illustrating tumour growth ( a ) and tumour volume ( b ). c The mRNA level of CD133 was determined by qPCR. d Tumour growth and volume were evaluated after treatment with DDP (3.0 mg kg −1 body weight; i.p., thrice) or PBS (pH 7.4; i.p., thrice). e Immunostaining images showing Notch2 expression. The mRNA levels of Notch2 and miR-181b were determined by qPCR. Mice were treated with A549/DDP sphere cells and then with DAPT or the control. f Images show tumour growth and tumour volume. g , h The mRNA levels of CD133 and Notch2 were measured by qPCR. i PARP and cleaved PARP levels were also ascertained by Western blotting after the indicated treatment. Data are presented as the mean ± SD. * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: We assessed the expression of the CD133 surface marker after transfection with miR-181b mimics (Invitrogen) or Notch2 siRNA (Santa Cruz Biotechnology).

Techniques: Over Expression, In Vivo, Transfection, Immunostaining, Expressing, Control, Western Blot

Journal: Stem Cell Research & Therapy

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

doi: 10.1186/s13287-018-1072-1

Figure Lengend Snippet: The clinical relevance of miR-181b and Notch2 in NSCLC. a miR-181b expression in all 8 NSCLC tumour specimens (T) compared with paired adjacent non-tumour (ANT) tissue as determined by qPCR. b Immunostaining images show Notch2 in tumour tissue and corresponding adjacent non-cancerous tissue. c miR-181b expression measured by qPCR in NSCLC patients at different disease stages. d A statistically significant inverse correlation between miR-181b and Notch2 mRNA levels in NSCLC tissues. e The expression of Notch2, CD133, and SOX2 in tumour tissues from 2 patients is shown by IHC. f , g OS and DFS curves for the high miR-181b expression group and the low miR-181b expression group, which were further analysed in different stages. h , i OS and DFS curves for the high-Notch2 expression group and the low-Notch2 expression group, which were further analysed in different stages. Data are presented as the mean ± SD. ** p < 0.01; *** p < 0.001

Article Snippet: We assessed the expression of the CD133 surface marker after transfection with miR-181b mimics (Invitrogen) or Notch2 siRNA (Santa Cruz Biotechnology).

Techniques: Expressing, Immunostaining

Journal: Stem Cell Research & Therapy

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

doi: 10.1186/s13287-018-1072-1

Figure Lengend Snippet: Association between miR181b expression and clinicopathological factors in patients with NSCLC

Article Snippet: We assessed the expression of the CD133 surface marker after transfection with miR-181b mimics (Invitrogen) or Notch2 siRNA (Santa Cruz Biotechnology).

Techniques: Expressing

Journal: Stem Cell Research & Therapy

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

doi: 10.1186/s13287-018-1072-1

Figure Lengend Snippet: Prognostic factors for lung SCC patients

Article Snippet: We assessed the expression of the CD133 surface marker after transfection with miR-181b mimics (Invitrogen) or Notch2 siRNA (Santa Cruz Biotechnology).

Techniques: Expressing