Journal: Biomedical Reports

Article Title: Anti-allergic effect of the naturally-occurring conjugated linolenic acid isomer, jacaric acid, on the activated human mast cell line-1

doi: 10.3892/br.2015.517

Figure Lengend Snippet: Jacaric acid modulates the expression levels of matrix metalloproteinase (MMP)-2, MMP-9 and tissue inhibitor of metalloproteinase-1 (TIMP-1) proteins in human mast cell line-1 (HMC-1) cells. HMC-1 cells were incubated with 1 (lane 2), 2 (lane 3) and 4 µM jacaric acid (lane 4) at 37°C for 72 h. Cells treated with 0.02% ethanol (lane 1) acted as the control. Subsequently, treated cells were further incubated with Iono and phorbol 12-myristate 13-acetate at 37°C for 6 h. (A) Protein expression levels of MMP-2, MMP-9 and TIMP-1 were assayed by western blotting with β-actin protein as an internal control. (B-D) The relative expression levels of MMP-9, MMP-2 and TIMP-1 proteins compared to β-actin were quantified. Results represent mean ± standard error. *P<0.05; **P<0.01; ***P<0.001.

Article Snippet: The membrane was first incubated with the following primary antibodies: Rabbit anti-MMP-2 (CST-4022S), anti-MMP-9 (CST-3852S), anti-tissue inhibitor of metalloproteinase-1 (TIMP-1) (CST-8946S) (Cell Signaling Technology, Inc., Danvers, MA, USA) and mouse anti-β-actin antibody (SC-A5316) (Santa Cruz Biotechnology, Inc., Dallas, TX, USA), followed by incubation with mouse IgG horseradish peroxidase-conjugated secondary antibody (GE-NA931) or rabbit IgG horseradish peroxidase-conjugated secondary antibody (GE-NA934) (GE Healthcare, Buckinghamshire, UK and finally developed with the enhanced chemiluminescence reagent (Santa Cruz Biotechnology, Inc.).

Techniques: Expressing, Incubation, Control, Western Blot