Fig. 4.

Lipopolyssacharide-induced inflammation profiles in Balb/c. Mice were injected intraperitoneally with LPS. After 20 h, the liver was collected and analyzed for Western blot, reverse transcriptase PCR, real time PCR, and immunohistochemistry. (A) Induction of pro-inflammatory cytokines, TNF-α, and stress inducible protein, heme oxygenase-1 (HO-1) in LPS-treated mouse. (B) Expression levels of TNF-α, IL-1β, and IL-6 mRNA were determined by RT-PCR in LPS injected mouse liver and observed by agarose-gel electrophoresis under UV illumination staining with ethidium bromide. (C) Liver lysates were prepared. Whole lysates (40 µg of protein) were separated by electrophoresis on SDS 10% polyacrylamide gels and then the protein was transferred onto nitrocellulose membrane. Each membrane was immunoblotted with antibody specific for phosphorylated or tatal JAK and STAT. (D) Levels of VCAM and ICAM mRNA were determined by semi-quantitative real time PCR in LPS injected mouse liver (Cont: ^*p<0.05, LPS: ^†p<0.01). (E) Liver from Balb/c mice injected with PBS or LPS were removed and embedded in paraffin, and then 4 µm sections were prepared. For immunohistochemistry studies, an immunohistochemistry kit was used, and all the procedures were performed according to the instructions of the manufacturer. The anti-TNF-α and anti-Tall like receptor (TLR) 4 were used. All results were representative of three separate experiments.