Files in this Data Supplement:

• Figure 1 - (Microsoft PowerPoint file) (71 KB). Generation of sPLA2-X-deficient mice. (A) Partial restriction map of the wild-type sPLA2-X allele (top), the targeting construct (middle), and the predicted homologous recombinant allele (bottom). Exon 1 including the ATG translation starting codon, Exon 2 and Exon 3 are indicated as a thick box. The thick bar indicates the position of the probe used for Southern blot hybridization. (B) Southern blot analysis of sPLA2-X locus. Genomic DNA was extracted from tail biopsies and digested with Pvu II. The blot was hybridized with probe. The wild-type allele gives a 10-kb fragment and the mutated allele gives a 4.8-kb fragment. +/+, wild-type mice; +/-, heterozygotes; -/-, homozygote. (C) RT-PCR analysis. cDNA synthesized from total RNA of stomach or colon, was used for PCR with specific primers for sPLA2-X.
• Figure II - (Microsoft PowerPoint file) (112 KB). Comparison of mRNA expression levels of various types of PLA2 between sPLA2-X^+/+ and sPLA2-X^-/- mice. Real-time quantitative PCR analysis of various types of PLA2 mRNA expression was performed in the left ventricle (LV) and the isolated peritoneal neutrophils from sPLA2-X^+/+(black bars) and sPLA2-X^-/- (white bars) mice. Upper panel shows agarose gel electrophoresis of the amplified PCR products at 25 cycles for cPLA2, sPLA2-V and PLA2-VI, and GAPDH and at 35 cycles for sPLA2-X from 0.1 Μg of total RNA. Lower panel summarizes the levels of mRNA expression normalized to GAPDH mRNA expression and expressed relative to the LV in sPLA2-X^+/+ mice (= 1). n = 5 ~ 7.
• Figure III - (Microsoft PowerPoint file) (101 KB).Comparison of mRNA expression levels between sPLA2-X^+/+ and sPLA2-X^-/- neutrophils. Expression levels of mRNA of NADPH oxidase p47phox and p67phox, elastase, CD11b/18, and cyclooxygenase-2 in the isolated neutrophils were measured with real-time quantitative PCR analysis of mRNA expression in sPLA2-X^+/+ (black bars) and sPLA2-X^-/- (white bars) mice. The upper panel shows agarose gel electrophoresis of the amplified PCR products at 25 cycles from 0.1 Μg of total RNA. The lower panel summarizes the levels of mRNA expression normalized to GAPDH mRNA expression and expressed relative to the sPLA2-X^+/+(=1) (n = 5~8 in each experiment).
• Methods and Tables - (Microsoft Word file) (117 KB).