Effects of phenylhydrazine treatment. (A) Plasma was extracted as described above from +/+ and −/− mice at 0, 24, and 96 hours after phenylhydrazine injections. One microliter of plasma was analyzed by Western blotting as described above. (B) RT-PCR using RNA from livers of +/+ and −/− mice at 0, 4, and 24 hours after phenylhydrazine injection of 2 mg/10 g body weight. RT-PCR was performed using Haptoglobin (Hp) and triose phosphate isomerase (TPI)-specific primers as described in Fig 1. The RT-PCR products were separated on a 5% polyacrylamide gel and exposed to autoradiography. The Hp signal was normalized against the TPI signal. Hp mRNA in livers of +/+ mice was elevated by 24 hours after phenylhydrazine injection. NoHp mRNA was observed in both livers and kidneys of −/− mice. (C) Mortality, hematocrit levels, and MDA/HNE levels of mice treated with phenylhydrazine. (D) Plasma AGP levels. Age-matched mice received either two subdermal injections of turpentine (Turp.), a single IP injection of PBS alone, or a single IP injection of the indicated concentrations of phenylhydrazine. Animals were bled before (0 hours, control) and 48 hours after injection. The concentration of AGP was determined by immunoelectrophoresis and the mean ± SD for the number of animals in each group shown.