Fig. 4.

ABP expression in the LNG of WT and Cyp2a5-null mice. A, quantitative RNA-PCR analysis of Abpa27 mRNA expression in B6 WT and Cyp2a5-null mice. LNGs from individual 2- to 3-month-old male mice were used for total RNA preparation. The relative levels of Abpa27 in LNG were determined. The results were corrected on the basis of the levels of GAPDH mRNA present in the same preparation. Data represent means ± S.D. (n = 4); **, P < 0.01, Student's t test, for comparisons between WT and Cyp2a5-null mice. B, immunoblot analysis of ABP protein levels in the LNG cytosol preparation from B6 WT and Cyp2a5-null mice. Cytosol fractions were prepared from pooled LNGs from 10 2- to 3-month-old male WT or Cyp2a5-null mice. Levels of GAPDH protein were also determined as a loading control. Samples (40 μg of protein per lane) were analyzed with use of either an anti-ABP antibody or an anti-GAPDH antibody. Densitometric analysis (not shown) indicated a ∼3-fold higher level of ABP expression in the Cyp2a5-null than in WT LNG. C, immunoblot analysis of ABP protein expression in subcellular fractions of the WT LNG. Pooled LNGs from 10 2- to 3-month-old male B6 WT mice were used for the preparation of tissue homogenate, postmitochondrial S9 fraction (S9), cytosol, and microsome fractions. For immunoblot analysis, 40 μg of protein were analyzed per lane.