Supplementary material for Oh et al. (2000) Proc. Natl. Acad. Sci. USA97 (6), 2626-2631.



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Fig. 6. Representative semi-log view of real-time amplification plots of β-actin (A), α 1 type IV collagen (Col4a1) (B), vascular endothelial growth factor (VEGF) (C), and tissue-type plasminogen activator (tPA) (D) during 40 PCR cycles. For each gene, several dilutions of wild-type (WT) cDNAs (10´, 1´, 0.1´, and 0.01´) were used to measure the relative amount of transcripts in homozygous (-/-; red), heterozygous (+/-; green), and WT (yellow) embryonic day (E)9.5 embryos. After initial normalization with β-actin primer, cDNA amount from each genotype that is equivalent to 1´ WT cDNA was used for the PCR amplification. (A) PCR products amplified by β-actin primers using cDNA from WT, +/-, and -/- E9.5 embryos were well aligned to 1´ cDNA from WT, verifying the normalization of the initial amount of total cDNA for the amplification. (B) The amplification plot indicates that Col4a1 transcripts were unaltered in activin receptor-like kinase (ALK)1+/- embryos. (C and D) The amplification plot of VEGF (C) and tPA (D) in -/- (red) was shifted to left (10´ WT), indicating elevated levels of VEGF and tPA transcripts in ALK1-/-, compared to wild-type and ALK1+/- embryos.