Figure 2. Aging associated adipose tissue inflammation is not correlated with the expression of TLR4 or its endogenous ligand, Fet A. Relative expression of Tlr4 gene in the adipose tissue of young or old mice at mRNA (A) and protein (B) levels. NF-κB activation was determined by western blotting using NF-κB-phospho-serine311 antibody (B). The relative density is expressed in (C). (D) Serum Fet A levels in young and old mice. (E) Relative mRNA expression of Fet A in the liver. mRNA expression of Fet A (F) and protein levels (G) in the adipose tissue of young and old mice. Data represented in bar diagrams are mean + SD value of relative mRNA expression from three independent experiments where total RNA was extracted from gonadal fat pads of young (n=5) and old (n=5) mice and used as a template for one-step qRT-PCR reaction. Protein expressions were determined by western blotting of adipose tissue lysates from young (n=5) and old (n=5) mice. Data presented here are representative image of three independent experiments. The significance levels *p<0.05, **p<0.001 were analyzed by unpaired Student’s t-test using means and SDs.