PCR identification of apo A-II-deficient mice,
Northern blot analysis of mouse liver RNA, and SDS/PAGE analysis of
mouse HDL. (A) The wild-type allele was identified by the
PCR primer pair MAII6u and MAII2l, which gave a 0.35-kb band. The
mutant allele was identified by the PCR primer pair MAII4l and Neo901,
which gave a 0.7-kb band. (B) RNA was isolated from mouse
liver. The Northern blot on the left was hybridized with mouse apo A-I
cDNA probe; the Northern blot on he right was hybridized with mouse apo
A-II cDNA probe. Apo A-II is completely absent from the apo A-II
knockout lane. The mRNA levels of heterozygous are only half of that of
controls. Apo A-I mRNA levels were unchanged among the three genotypes.
+/+, +/−, and −/− represent control littermates,
heterozygous-deficient mice, and homozygous apo A-II-deficient mice,
respectively. (C) HDL was isolated by sequential
ultracentrifugation and HDL from equal amounts of plasma was subjected
onto SDS/PAGE (10–25% gradient). The gel was stained with
Coomassie blue R-250. There was no detectable apo A-II in the
homozygous-deficient mouse HDL.