Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Anti-inflammatory effects of mitogen-activated protein kinase kinase inhibitor U0126 in an asthma mouse model.

doi: 10.4049/jimmunol.172.11.7053

Figure Lengend Snippet: FIGURE 7. Effects of U0126 on ERK1/2 phosphorylation in lung tis- sue. Western blot analysis of OVA-induced ERK1/2 tyrosine phosphory- lation in lung fragments isolated from mice treated with PEG and 30 mg/kg U0126. Lung fragments were homogenized in ice-cold lysis buffer. Pro- teins (20 g/lane) were separated by SDS-PAGE and probed with anti-pan ERK and anti-phospho-ERK Abs. The blot was developed by nitroblue tetrazolium and 5-bromo-4-chloro-3-indolyl phosphate, and analyzed using Gel-Pro imaging software. Results are expressed as fold increase of basal ERK level in naive mice (A and B). Basal ERK level is given a value of 1. Values shown are the mean SEM of three separate experiments. , Sig- nificant difference from naive, and , significant difference from PEG control, p 0.05. N, naive; O, OVA; P, PEG; U, U0126.

Article Snippet: The membrane was probed with anti-pan ERK and anti-phospho-ERK (Thr202/Tyr204) Abs (Cell Signaling Technology, Beverly, MA), followed by alkaline phosphatase-conjugated secondary Ab, and visualized using chromogenic substrates nitroblue tetrazolium and 5-bromo-4-chloro-3-indolyl phosphate (Life Technologies, Gaithersburg, MD).

Techniques: Phospho-proteomics, Western Blot, Isolation, Lysis, SDS Page, Imaging, Software, Control