human netrin 4 elisa kit  (Novus Biologicals)

Journal: International Journal of Clinical and Experimental Pathology

Article Title: miR-17-5p promotes migration and invasion in breast cancer cells by repressing netrin 4

doi:

Figure Lengend Snippet: Markedly negative correlation between miR-17-5p and NTN4 mRNA in BC lesions in a database analysis. A. An analysis of NTN4 mRNA expression in 113 normal breast tissues and 1109 BC tissues in the TCGA database. B. miR-17-5p expression in 104 normal breast specimens and 1103 BC samples from the data in TCGA. C. Serum miR-17-5p expression in GSE73002, including 2686 noncancer controls and 1280 BC samples, ***P < 0.001. D, E. Correlations between miR-17-5p and NTN4 mRNA in TCGA (1066 pairs of samples, P < 0.0001) and GSE22220 (207 pairs of tissues, P = 0.010). F, G. Survival analysis of miR-17-5p and NTN4 in BC with a Kaplan-Meier plotter. H, I. Progression-free survival of miR-17-5p and NTN4 in GSE22220.

Article Snippet: NTN4 protein concentration was measured with an ELISA kit (CSB-E11900h; CUSABIO, China, https://www.cusabio.com/), according to manufacturer’s instructions, as described previously [ 9 ].

Techniques: Expressing

Journal: International Journal of Clinical and Experimental Pathology

Article Title: miR-17-5p promotes migration and invasion in breast cancer cells by repressing netrin 4

doi:

Figure Lengend Snippet: Validation of miR-17-5p transfection. A. miR-17-5p and NTN4 mRNA expression in four BC cell lines, determined with RT-qPCR. B, C. Fold change in miR-17-5p expression after cell transfection with miR-17 mimic, miR-17 inhibitor, or their matching controls. ‘Mimic NC’ denotes mimic-transfected negative control, and ‘inhibitor NC’ denotes inhibitor-transfected negative control. D, E. Influence of miR-17-5p on colony formation. D. miR-17-5p upregulation in Hs578T cells promoted colony formation. E. Reduced miR-17-5p in MCF-7 cells suppressed colony formation. Colony numbers are quantified in the right bar graphs. *P < 0.05, ***P < 0.001.

Article Snippet: NTN4 protein concentration was measured with an ELISA kit (CSB-E11900h; CUSABIO, China, https://www.cusabio.com/), according to manufacturer’s instructions, as described previously [ 9 ].

Techniques: Biomarker Discovery, Transfection, Expressing, Quantitative RT-PCR, Negative Control

Journal: International Journal of Clinical and Experimental Pathology

Article Title: miR-17-5p promotes migration and invasion in breast cancer cells by repressing netrin 4

doi:

Figure Lengend Snippet: miR-17-5p directly binds to NTN4 mRNA. A. Expression of NTN4 mRNA (left panel) and protein (right panel) changed after the transfection of BC cells with miR-17 mimic or miR-17 inhibitor. B. Binding site of miR-17-5p within NTN4 mRNA predicted with TargetScan and miRDB. C. h-NTN4-WT and h-NTN4-MUT were constructed in the pmiR-RB-REPORT vector, with wild-type and mutated miR-17-5p binding sites inserted, respectively, downstream from the luciferase reporter. D. In the luciferase reporter assay, miR-17-5p significantly reduced the luciferase activity expressed from h-NTN4-WT (P < 0.05) but not that from h-NTN4-MUT (P > 0.05). *P < 0.05, **P < 0.01.

Article Snippet: NTN4 protein concentration was measured with an ELISA kit (CSB-E11900h; CUSABIO, China, https://www.cusabio.com/), according to manufacturer’s instructions, as described previously [ 9 ].

Techniques: Expressing, Transfection, Binding Assay, Construct, Plasmid Preparation, Luciferase, Reporter Assay, Activity Assay

Journal: Oncology reports

Article Title: NTN4 is associated with breast cancer metastasis via regulation of EMT-related biomarkers.

doi: 10.3892/or.2016.5239

Figure Lengend Snippet: Figure 3. Overexpression of NTN4 inhibits the migration and invasion of MDA-MB-231 cells. (A and B) The overexpression of NTN4 in MDA-MB-231 cells by transient transfection of pcDNA3.1-NTN4 and pcDNA3.1 was confirmed with RT-qPCR (A) and ELISA (B) after transfection for 48 h. (C) NTN4 overex- pression reduced the wound healing rate of MDA-MB-231 cells. (D and E) Overexpression of NTN4 inhibited the migration and invasion of MDA-MB-231 cells. Cells transfected with pcDNA3.1 or pcDNA3.1-NTN4 were subjected to the migration and invasion assays, and the migrated and invaded cells were fixed at 24 and 48 h, respectively. Magnification, x400. Mean ± SD, n≥3. *p<0.05, **p<0.01.

Article Snippet: NTN4 concentration of cell supernatant was measured using an enzyme linked immunosorbent assay (ELISA) kit (CSB-E11900h; CusaBio, Wuhan, China).

Techniques: Over Expression, Migration, Transfection, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Oncology reports

Article Title: NTN4 is associated with breast cancer metastasis via regulation of EMT-related biomarkers.

doi: 10.3892/or.2016.5239

Figure Lengend Snippet: Figure 4. Silencing of NTN4 promotes migration and invasion of Hs578T cells. (A) qRT-PCR and (B) ELISA revealed that siRNA knockdown of NTN4 reduced mRNA and protein levels of NTN4 in Hs578T cells after transfection for 48 h. (C) NTN4 knockdown accelerated the wound healing rate in Hs578T cells. (D and E) NTN4 silencing enhanced cell migration and invasion activity. Cells transfected with siRNA NC or NTN4 siRNA were subjected to the migra- tion and invasion assays, and the migrated and invaded cells were fixed at 24 and 48 h, respectively. Magnification, x400. Mean ± SD, n≥3. *p<0.05, **p<0.01.

Article Snippet: NTN4 concentration of cell supernatant was measured using an enzyme linked immunosorbent assay (ELISA) kit (CSB-E11900h; CusaBio, Wuhan, China).

Techniques: Migration, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Knockdown, Transfection, Activity Assay