human mmp 15  (R&D Systems)

Journal: Molecular Cancer

Article Title: Tumor cell invasion of collagen matrices requires coordinate lipid agonist-induced G-protein and membrane-type matrix metalloproteinase-1-dependent signaling

doi: 10.1186/1476-4598-5-69

Figure Lengend Snippet: HEK293 cells transfected with MT1-MMP cDNA invade 3D collagen gels in response to LPA . (A) Tumor cell lysates were prepared for Western blot analysis. Lysates were probed for MT1-MMP to assess protein expression in the four tumor cell lines. Lysates were probed for Actin as a loading control. (B) HEK293 cells were transfected with the pAdTrack-CMV plasmid as a control, or plasmids encoding MT1-MMP, MT2-MMP, or MT3-MMP cDNA 24 hours prior to placement in invasion assays. Cells were allowed to invade 2.0 mg/ml collagen gels in the presence or absence of 1 μM LPA. Data are expressed as mean numbers of invading cells per HPF (20×) (± S.D.) from a minimum of 20 fields. (C) Lysates from HEK293 cells transfected with cDNAs encoding the designated genes were prepared for Western blot analysis and probed for GFP, MT1-MMP, MT2-MMP, MT3-MMP, or Actin as a loading control. TRK = pAdTrack-CMV, MT1 = MT1-MMP, MT2 = MT2-MMP, MT3 = MT3-MMP.

Article Snippet: A plasmid encoding human MT2-MMP (TC118648) was purchased from Origene (Rockville, MD).

Techniques: Transfection, Western Blot, Expressing, Plasmid Preparation

Journal: International journal of molecular sciences

Article Title: Silencing PROK2 Inhibits Invasion of Human Cervical Cancer Cells by Targeting MMP15 Expression.

doi: 10.3390/ijms21176391

Figure Lengend Snippet: Figure 3. Effect of knockdown PROK2 on MMP15 expression and cell invasion in human cervical cancer HeLa cells. (A) Human HeLa cells were transfected with or without PROK2 shRNA, then followed by measuring the capacity of cell migration and invasion. (B,C) The protein and mRNA expression of MMP15 were inhibited by shPROK2-HeLa cells were measured by western blotting and R-qPCR assay. (D) Validation of MMP15 gene expression in matched cervical cancer tissues and adjacent noncancerous cervical tissues from the GEPIA databases. T: cervical tumour tissue (n = 306); N: normal cervical tissue (n = 13), * p < 0.05 versus normal cervical tissue. (E) Overall survival rate (OS) in patients with high or low MMP15 expression. The red line indicates high expression, and black line indicates low expression. (F) MMP15 expression was correlated with PROK2 expression in human cervical cancer patients. ** p < 0.01 versus shLuc cells.

Article Snippet: Antibodies against MMP15 (130522, 1:500) was obtained from Novus Biologicals (Briarwood, CO, USA) and horseradish peroxidase (HRP)-conjugated anti-mouse (AP124P, 1:10,000) were purchased from Merck Millipore (Burlington, MA, USA).

Techniques: Knockdown, Expressing, Transfection, shRNA, Migration, Western Blot, Biomarker Discovery, Gene Expression

Journal: International journal of molecular sciences

Article Title: Silencing PROK2 Inhibits Invasion of Human Cervical Cancer Cells by Targeting MMP15 Expression.

doi: 10.3390/ijms21176391

Figure Lengend Snippet: Figure 4. MMP15 involved in PROK2 regulates cell migration and invasion in human cervical cancer HeLa cells. Using transfected with Neo or PROK2 overexpression plasmid in shLuc- or shPROK2-HeLa cells for 48 h. (A) The protein expression of MMP15 and PROK2 were measured by the western blotting. β-actin as a protein loading control. (B) The MMP15 and PROK2 mRNA expression were detected by RT-qPCR assay. GAPDH as a mRNA loading control. (C) In vitro migration and invasion assay was conducted to measures the cell migration and invasion numbers. ** p < 0.01 versus shLuc cells; # p < 0.05 verus shPROK2 cells.

Article Snippet: Antibodies against MMP15 (130522, 1:500) was obtained from Novus Biologicals (Briarwood, CO, USA) and horseradish peroxidase (HRP)-conjugated anti-mouse (AP124P, 1:10,000) were purchased from Merck Millipore (Burlington, MA, USA).

Techniques: Migration, Transfection, Over Expression, Plasmid Preparation, Expressing, Western Blot, Control, Quantitative RT-PCR, In Vitro, Invasion Assay