incubation by elisa  (R&D Systems)

Journal: Cell Death & Disease

Article Title: Circular RNA circ_001621 promotes osteosarcoma cells proliferation and migration by sponging miR-578 and regulating VEGF expression

doi: 10.1038/s41419-019-2204-y

Figure Lengend Snippet: a Prediction of putative-binding sites between miR-578 and VEGF by miRDB. b Negative correlation between miR-578 expression and VEGF. c Luciferase reporter assay indicated that VEGF DEL abrogated the inhibition of VEGF promoter activity. ** P < 0.01, vs control. d–g VEGF expression in cells expressing control, miR-578 or miR-578 inhibitor was detected by western blot and real-time PCR, respectively. Results represented the mean ± s.d. of three independent experiments. ** P < 0.01, vs control. miR-578 in, miR-578 inhibitor.

Article Snippet: Then membranes were blocked with 5% blocking solution for 1 h, followed by incubation with VEGF (19003-1-AP), cyclin-dependent kinase 4 (CDK4) (11026-1-AP), and matrix metallopeptidase 9 (MMP9) (10375-2-AP) antibodies (Proteintech Group, Inc, Rosemont, IL, USA) overnight at 4 °C.

Techniques: Binding Assay, Expressing, Luciferase, Reporter Assay, Inhibition, Activity Assay, Control, Western Blot, Real-time Polymerase Chain Reaction

Journal: Cell Death & Disease

Article Title: Circular RNA circ_001621 promotes osteosarcoma cells proliferation and migration by sponging miR-578 and regulating VEGF expression

doi: 10.1038/s41419-019-2204-y

Figure Lengend Snippet: a Proliferation in cells expressing control, circ_001621 or combination was detected by the MTT assay. b Migration of MG63 cells expressing control, circ_001621 or combination was detected by the transwell assay. The indicated genes expression in MG63 cells were detected by western blot ( c ) and real-time PCR ( d ), separately. Results represented the mean ± s.d. of three independent experiments. ** P < 0.01, vs control, ## P < 0.01, vs circ_001621. e Immunofluorescent stain of VEGF in the indicated MG63 cells. Green, VEGF; Blue, DAPI.

Article Snippet: Then membranes were blocked with 5% blocking solution for 1 h, followed by incubation with VEGF (19003-1-AP), cyclin-dependent kinase 4 (CDK4) (11026-1-AP), and matrix metallopeptidase 9 (MMP9) (10375-2-AP) antibodies (Proteintech Group, Inc, Rosemont, IL, USA) overnight at 4 °C.

Techniques: Expressing, Control, MTT Assay, Migration, Transwell Assay, Western Blot, Real-time Polymerase Chain Reaction, Staining

Journal: Cell Death & Disease

Article Title: Circular RNA circ_001621 promotes osteosarcoma cells proliferation and migration by sponging miR-578 and regulating VEGF expression

doi: 10.1038/s41419-019-2204-y

Figure Lengend Snippet: The lung ( a ) and liver metastatic foci ( b ) were validated by hematoxylin and eosin staining. The number of metastatic foci in each mouse was shown in the right panel. con control, circ circ_001621, miR miR-578. ** P < 0.01, vs control; ## P < 0.01, vs circ_001621. c Immunohistochemistry of VEGF expression in lung metastasis. The indicated genes expression was measured using western blot ( d ) and real-time PCR ( e ), respectively. Results represented the mean ± s.d. of three independent experiments. ** P < 0.01, vs control; ns, P > 0.05, vs circ_001621; ## P < 0.01, vs circ_001621.

Article Snippet: Then membranes were blocked with 5% blocking solution for 1 h, followed by incubation with VEGF (19003-1-AP), cyclin-dependent kinase 4 (CDK4) (11026-1-AP), and matrix metallopeptidase 9 (MMP9) (10375-2-AP) antibodies (Proteintech Group, Inc, Rosemont, IL, USA) overnight at 4 °C.

Techniques: Staining, Control, Immunohistochemistry, Expressing, Western Blot, Real-time Polymerase Chain Reaction