Journal: Experimental and Therapeutic Medicine

Article Title: In vitro and in vivo effects of AVA4746, a novel competitive antagonist of the ligand binding of VLA-4, in B-cell acute lymphoblastic leukemia

doi: 10.3892/etm.2021.10969

Figure Lengend Snippet: AVA4746 decreases cell surface and total integrin α4 expression. MFI representing the expression levels of integrin α4 on the (A) LAX7R, (B) LAX7, (C) LAX53, (D) LAX56 and (E) REH cell surfaces 24 and 96 h after treatment with 1, 5 and 25 µM AVA4746, with 0 representing the DMSO control. * P<0.05 vs. 0. Experiments were performed in triplicates. (F) B-ALL cell lines LAX7R, LAX7, LAX53, LAX56 and REH were treated for 24 and 96 h with 1, 5 and 25 µM AVA4746, with 0 representing the DMSO control. Following 24 and 96 h treatment with AVA4746, the integrin α4 protein levels at 140 kDa in LAX7R, LAX7, LAX53, LAX56 and REH cells were analyzed by western blotting, with β-actin being used as lthe oading control. MFI, mean fluorescence intensity; α4, α4 integrin.

Article Snippet: The following primary antibodies were incubated overnight at 4˚C: Anti-integrin α4 (#4600, 1:1,000; Cell Signaling Technology, Inc.), anti- phosphorylated (p-)-AKTSer473 (#9271, 1:1000; Cell Signaling Technology, Inc.), anti-AKT (#9272, 1:1,000; Cell Signaling Technology, Inc.), anti-phosphotyrosine (#05-321MG, 1:1000 dilution, EMD Millipore) and anti-β-actin (#sc-47778, 1:2000 dilution, Santa Cruz Biotechnology, Inc.).

Techniques: Expressing, Western Blot, Fluorescence

Journal: Experimental and Therapeutic Medicine

Article Title: In vitro and in vivo effects of AVA4746, a novel competitive antagonist of the ligand binding of VLA-4, in B-cell acute lymphoblastic leukemia

doi: 10.3892/etm.2021.10969

Figure Lengend Snippet: AVA4746 attenuates tube formation by HUVECs. HUVECs treated with either 0, 5 or 25 µM AVA4746 for 30 min were seeded into Matrigel-coated plates for 4-6 h. (A) Representative flow cytometry dot plot of human integrin α4 and β1 expression in HUVECs before seeding into the Matriel wells. (B) Representative images of the HUVECs are shown. (C) Tube formation was assessed by analyzing the number of nodes (pixels with ≥ three neighboring elements corresponding to a bifurcation), segments (elements delimited by two junctions), meshes (areas enclosed by segments or master segments and made by tube-like structures) and total area, before being subsequently quantified. Images were analyzed with using the ImageJ software and the ‘angiogenesis analyzer’ plugin. * P<0.05, ** P<0.01 and *** P<0.001. HUVEC, Human umbilical vein endothelial cells; α4, α4 intergrin; β1, β1 integrin; Nb, number.

Article Snippet: The following primary antibodies were incubated overnight at 4˚C: Anti-integrin α4 (#4600, 1:1,000; Cell Signaling Technology, Inc.), anti- phosphorylated (p-)-AKTSer473 (#9271, 1:1000; Cell Signaling Technology, Inc.), anti-AKT (#9272, 1:1,000; Cell Signaling Technology, Inc.), anti-phosphotyrosine (#05-321MG, 1:1000 dilution, EMD Millipore) and anti-β-actin (#sc-47778, 1:2000 dilution, Santa Cruz Biotechnology, Inc.).

Techniques: Flow Cytometry, Expressing, Software