Journal: Molecular Medicine Reports
Article Title: Roles of lipoxin A4 receptor activation and anti-interleukin-1β antibody on the toll-like receptor 2/mycloid differentiation factor 88/nuclear factor-κB pathway in airway inflammation induced by ovalbumin
doi: 10.3892/mmr.2015.3443
Figure Lengend Snippet: Activation of NF-κB assessed using western blot analysis in leukocytes exposed to IL-1β. The cultured leukocytes were obtained from a normal control mouse and stimulated with IL-1β (10 ng/ml) for 1 h with or without pre-treatment of BML-111 (1 mM) for 30 min, BOC1 (100 μ M) for 30 min, TLR2Ab (1 μ g/ml) for 1 h, goat IgG (1 μ g/ml) for 1 h, ST2825 (20 μ M) for 1 h or TPCA-1 (10 μ M) for 1 h. The western blot is representative of five independent experiments. GAPDH served as a loading control of cytosolic proteins. Histone 3 protein served as a loading control of nuclear proteins. Semiquantitative analysis was performed using UVP-gel densitometry. Arbitrary unit = (A IκBα /A GAPDH ) ×100%, or (A cytosolic p65 /A GAPDH ) ×100%, or (A nuclear p65 /A histone 3 ) ×100%. Values are expressed as the mean ± standard deviation of five independent experiments. * P<0.05, as compared to the arbitrary units of the same protein in the cells without treatment. IL, interleukin; IgG, immunoglobulin G; TPCA-1,10, thiophene-3-carbox-amide 1; TLR2Ab, toll-like receptor 2-neutralizing antibody; BOC1, N -t-Boc-Phe-Leu-Phe-Leu-Phe; NF-κB, nuclear factor-κB; IκBα, inhibitor of kappa B alpha.
Article Snippet: The blots were incubated with primary rabbit anti-mouse polyclonal antibodies against TLR2 (cat. no. sc-10739), MyD88 (cat. no. sc-11356) or β-actin (cat. no. sc-130656; Santa Cruz Biotechnology, Inc.) at 1:2,000 dilution overnight followed by incubation for 2 h with an HRP-conjugated goat anti-rabbit IgG antibody (Jackson, West Grove, PA, USA) at 1:5,000 dilution.
Techniques: Activation Assay, Western Blot, Cell Culture, Control, Standard Deviation