2-DE analysis of normal and infected hamsters urine. Urinary protein patterns of hamsters were compared by 2-DE analysis; (A, C) before infection; (B, D) 7–8 days post-infection. The urine of three hamsters was mixed for each infection period. The total protein content of each sample was 20 μg. Each pattern of urinary protein was separated by pI (4–7), 12.5% acrylamide gel, and subsequently silver staining (A, B), or immunoblotting with anti-L. interrogans pAb was done (C, D). Arrows (D) show spots of 60 kDa that reacted with the polyclonal antibody at 7–8 days post-infection. Each experiment was repeated three times, and the representative data are shown in this figure.